碱性磷酸酶显色试剂盒(40x)

碱性磷酸酶显色试剂盒(40x)

有货

碱性磷酸酶显色试剂盒(40x)

品牌:Jinpan
BCIP/NBT Kit(40x)

MSDS

质检证书(CoA)

相似产品

货号 (SKU) 包装规格 是否现货 价格 数量
B273090-40ml 40ml 期货 碱性磷酸酶显色试剂盒(40x)  

基本信息

产品名称 碱性磷酸酶显色试剂盒(40x)
英文名称 BCIP/NBT Kit(40x)
运输条件 冰袋运输

一般描述

产品简介

BCIP (5-Bromo-4-chloro-3-indolyl phosphate) 5-溴-4-氯-3-吲哚-磷酸盐+NBT (四唑硝基蓝) 是碱性磷酸酶(AP)最佳的底物组合之一。在碱性磷酸酶的催化下,BCIP会被水解而产生强反应性的产物,该产物与NBT发生反应,形成不溶性的深蓝色至蓝紫色化合物。该试剂盒可用于AP系统的IHC 和Western Blot 实验的酶促显色。在AP催化下,在组织切片或印迹膜上结合了AP偶联物的地方产生深蓝色沉淀,可根据颜色反应来确定目的蛋白的位置及表达情况。

产品组分

40×BCIP:1 ml

40×NBT:1 ml

BCIP/NBT Buffer:40 ml

注意事项

1.工作液应现配现用,配制好的工作液1小时内有效。

2.工作液用量必需充足,保证完全覆盖组织片或印迹膜。 

3.为获得最佳实验结果,请务必优化实验条件。

4.NBT有毒,使用时请采取必要的防护措施。

5.本产品仅用于科研,不能用于人体实验或人体治疗。

使用方法

1.BCIP/NBT显色工作液配制:

根据需要量,将40×BCIP、40×NBT和BCIP/NBT Buffer以1:1:38的体积比混匀后,即为BCIP/NBT显色工作液。

2.显色:

1)印迹膜显色:将配制好的工作液滴加在印迹膜上(或将印迹膜倾入到BCIP/NBT显色工作液中),室温避光孵育3-10分钟。显色完毕后,将膜浸入水中,终止反应。

2)组织切片或细胞爬片显色:滴加适量的BCIP/NBT显色工作液于需要显色的组织切片或细胞爬片上,室温避光孵育3-10分钟。显微镜下观察控制显色时间,当达到最佳显色效果后,自来水冲洗终止显色。显色后的切片经复染、脱水透明,封片后可长期保存。

Product Introduction

BCIP (5-Bromo-4-chloro-3-indolyl phosphate) 5-bromo-4-chloro-3-indolyl-phosphate + NBT (tetrazolium nitro blue) is the best substrate for alkaline phosphatase (AP) One of the combination. Under the catalysis of alkaline phosphatase, BCIP will be hydrolyzed to produce a highly reactive product, which reacts with NBT to form an insoluble dark blue to blue-violet compound. This kit can be used for the enzymatic color development of IHC and Western Blot experiments of the AP system. Under AP catalysis, a dark blue precipitate is produced where AP conjugates are combined on tissue sections or blotting membranes. The location and expression of the target protein can be determined based on the color reaction.

Product Components

40×BCIP: 1 ml

40×NBT: 1 ml

BCIP/NBT Buffer: 40 ml

Precautions

1. The working fluid should be prepared for immediate use, and the prepared working fluid will be effective within 1 hour.

2. The amount of working fluid must be sufficient to ensure complete coverage of the tissue sheet or blotting membrane. To

3. In order to obtain the best experimental results, be sure to optimize the experimental conditions.

4. NBT is poisonous, please take necessary protective measures when using it.

5. This product is only used for scientific research, not for human experiments or human treatment.

Instructions

1. BCIP/NBT color developing working solution preparation:

According to the required amount, mix 40×BCIP, 40×NBT and BCIP/NBT Buffer in a volume ratio of 1:1:38 to form the BCIP/NBT color developing working solution.

2. Color rendering:

1) Blotting membrane color development: Drop the prepared working solution on the blotting membrane (or pour the blotting membrane into the BCIP/NBT color developing working solution), and incubate for 3-10 minutes at room temperature and dark. After the color development is completed, the film is immersed in water to terminate the reaction.

2) Color development of tissue sections or cell slides: Drop an appropriate amount of BCIP/NBT color developing working solution on the tissue sections or cell slides that need color development, and incubate at room temperature for 3-10 minutes in the dark. Observe under the microscope to control the color development time. When the best color development effect is reached, rinse with tap water to stop the color development. After color development, the slices are counter-stained, dehydrated and transparent, and can be stored for a long time after mounting.

相关属性

储存温度 2-8°C储存,避光,干燥
品牌 Jinpan

台盼蓝染色试剂盒

台盼蓝染色试剂盒

0.40%

有货

台盼蓝染色试剂盒

CAS编号 72-57-1 | 品牌:Jinpan
Trypan Blue Dyeing Kit

MSDS

质检证书(CoA)

相似产品

  • 分子式 C34H24N6O14S4Na4
  • 分子量960.81
  • Beilstein号 4360496
  • EC号 200-786-7
  • PubChem编号 6296

货号 (SKU) 包装规格 是否现货 价格 数量
T293438-50ml 50ml 期货 台盼蓝染色试剂盒  

基本信息

产品名称 台盼蓝染色试剂盒
英文名称 Trypan Blue Dyeing Kit
规格或纯度 0.40%
运输条件 常规运输

一般描述

产品介绍

台盼蓝染色试剂盒(Trypan Blue Dyeing Kit)用于细胞培养实验中活细胞,死细胞计数以及细胞存活率测定实验。台盼蓝是细胞活性染料,常用于检测细胞膜的完整性与细胞的存活率,是组织和细胞培养中最常用的死细胞鉴定染色方法之一。正常的活细胞细胞膜结构完整,能够排斥台盼蓝,细胞不会被染成蓝色;而丧失活性或细胞膜不完整的细胞可被台盼蓝染成蓝色。通常认为细胞膜完整性丧失,即可认为细胞已经死亡。台盼蓝可被巨噬细胞吞噬,故可用于吞噬细胞的活体染色剂。凋亡小体也有台盼蓝拒染现象。台盼蓝染色后,通过显微镜下直接计数货显微镜下拍照后计数,就可以对细胞存活率进行比较精确的定量。染色时间只需3-5分钟,操作简单。

T293438 由50 ml染液(0.4%)和100 ml重悬液组成(注:更多重悬液可用产品 P301984 替代)

使用方法:

1.制备细胞悬液:用胰酶和/或EDTA消化贴壁细胞,悬浮细胞可直接收集,收集细胞时用1000-2000rpm离心1分钟,弃上清,用细胞重悬液制备单细胞悬液,并作适当稀释

2.染色:细胞悬液与台盼蓝溶液以9:1混合均匀,染色3分钟(染色3分钟时间已经足够,染色时间可以更长一些,但不宜超过10分钟。

3.细胞计数:吸取少量经过染色的细胞,用血细胞计数板计数。死细胞着蓝色并膨大,无光泽;活细胞不着色并保持正常形态,有光泽。(通常要比较精确的进行定量,每个细胞样品至少数500个细胞。

4.统计细胞活力:细胞存活率(%)=活细胞总数/(活细胞总数+死细胞总数)x100%。
应用范围:
适用于一般生物学及医学研究。

相关属性

CAS编号 72-57-1
熔点 >300°C
RTECS QJ6475000
分子量 960.81
分子式 C34H24N6O14S4Na4
EC号 200-786-7
品牌 Jinpan
Smiles [Na+].[Na+].[Na+].[Na+].Cc1cc(ccc1/N=Nc1c(O)c2c(N)cc(cc2cc1S(=O)(=O)[O-])S(=O)(=O)[O-])c1ccc(/N=Nc2c(O)c3c(N)cc(cc3cc2S(=O)(=O)[O-])S(=O)(=O)[O-])c(C)c1;Cc1cc(ccc1/N=N/c1c(O)c2c(N)cc(cc2cc1S(=O)(=O)O)S(=O)(=O)O)c1ccc(/N=N/c2c(O)c3c(N)cc(cc3cc2S(=O)(=O)O)S
PubChem CID 6296

ROS 活性氧检测试剂盒

ROS 活性氧检测试剂盒

有货

ROS 活性氧检测试剂盒

品牌:Jinpan
Reactive Oxygen Species Assay Kit

MSDS

质检证书(CoA)

相似产品

货号 (SKU) 包装规格 是否现货 价格 数量
R272916-1000T 1000T 现货 ROS 活性氧检测试剂盒  

基本信息

产品名称 ROS 活性氧检测试剂盒
英文名称 Reactive Oxygen Species Assay Kit
运输条件 超低温冰袋运输

一般描述

产品组成

 

产品名称

包装

A液

H2DCFH-DA(10mM)

0.1mL

B液

活性氧阳性对照(Rosup,50mg/mL)

1mL


产品简介

活性氧检测试剂盒(Reactive Oxygen Species Assay Kit)是一种利用荧光探针H2DCFH-DA进行活性氧检测的试剂盒。H2DCFH-DA本身没有荧光,可以自由穿过细胞膜,进入细胞内后,可以被细胞内的酯酶水解生成DCFH。而DCFH不能通透细胞膜,从而使探针很容易被装载到细胞内。细胞内的活性氧可以氧化无荧光的DCFH生成有荧光的DCF。检测DCF的荧光就可以知道细胞内活性氧的水平。根据活细胞中荧光的产生,可以判断细胞活性氧的含量和变化。用流式细胞仪或荧光显微镜可直接观察,是一种经典的组织或活细胞中活性氧检测方法。本试剂盒提供了活性氧阳性对照试剂Rosup,以便于活性氧的检测。Rosup是一种混合物,浓度为50mg/mL。Rosup 为活性氧阳性诱导药物,根据其荧光信号强度,可分析活性氧的真正水平。

本试剂盒本底低,灵敏度高,线性范围宽,使用方便。本试剂盒可以测定1000个样品1000T(96 孔板)。

一套包含96孔板,可使用1000次。
对光敏感,注意避光保存

注意事项

1. 探针装载后,一定要洗净残余的未进入细胞内的探针,否则会导致背景较高。

2. 阳性对照Rosup 一般使用浓度为100 μM (推荐浓度100-400μM,具体依细胞类型而定)。通常刺激后0.5-4h 可观察到显著的活性氧水平升高。对于不同的细胞,活性氧阳性对照的效果可能有较大的差别。如果在刺激后30min 内观察不到活性氧的升高,可延长诱导时间或适当提高活性氧阳性对照的浓度。如果活性氧升高得过快,可缩短诱导时间或适当降低活性氧阳性对照的浓度。

3. 对于某些特别的细胞,实验过程中如果发现没有刺激的阴性对照细胞荧光也比较强, 可以按照1:2000-1:5000 稀释DCFH-DA , 使DCFH-DA 的终浓度为2-5 μM。探针装载的时间也可以根据情况在15-60 min 内适当进行调整。

4. 活性氧阳性对照(Rosup) 仅仅用于作为阳性对照的样品,并不是在每个样品中都需加入活性氧阳性对照。

5. 探针装载完毕并洗净残余探针后,可以进行激发波长的扫描和发射波长的扫描,以确认探针的装载情况是否良好。DCF的激发光谱和发射光谱请参考上图。

6. 尽量缩短探针装载后到测定所用的时间(刺激时间除外),以减少各种可能的误差。

7. 为了您的安全和健康,请穿实验服并戴一次性手套操作。

8. 定量的话要作标准曲线吧。先做一个不同浓度H2O2氧化DCFA荧光值,做一条标准曲线,X轴为H2O2浓度,y轴是荧光值,得出一个方程,在看你样品的荧光值即Y值是多少,对应的X值就是。

9. 有的细胞装载探针后细胞容易漂起来,洗细胞时实验组会吸走一部分细胞。所以种细胞时细胞量增加一倍,这样细胞紧密连接,贴壁比较牢,实验组的荧光值就高了。另外的H2DCFDA很敏感,工作液浓度要低一些,1-2μM就够啦,浓度太高容易有非特异性染色。这个探针很不稳定,一旦氧化了本底荧光值就会升高,最好工作液现用现配。

使用说明

1. 装载ROS 探针

1.1 原位装载探针(仅适用于贴壁细胞)

a) 细胞准备:检测前一天进行细胞铺板,确保检测时细胞数量小于5×10^5/ml。

b) 药物诱导:去除细胞培养液,加入无血清培养基稀释的药物处理,于37℃细胞培养箱内避光孵育,实际诱导时间由药物特性和细胞类型决定。

c) (可选)阳性对照:先用无血清培养基等稀释阳性对照(Rosup, 100 mM)到常用工作浓度100 μM,加入细胞,37℃避光孵育0.5~4 h,以提高活性氧水平,不同细胞类型存在差异。例如:HeLa 细胞需孵育30-60 min,MRC5 人胚胎成纤维细胞则需孵育90 min。

d) ROS 探针准备:探针装载前按照1:1000 用无血清培养液稀释DCFH-DA,使其终浓度为10 μM。

e) ROS 探针装载:吸除处理药物,加入适当体积稀释好的DCFH-DA 工作液。加入的体积需充分盖住细胞。例如:6孔板通常不少于1000 μL,对于96 孔板通常不少于100 μL。37℃细胞培养箱内避光孵育30 min。

f) 细胞清洗:用无血清培养液洗涤细胞1~2 次,以充分去除未进入细胞内的DCFH-DA。

1.2 收集细胞后装载探针(适用于贴壁细胞和悬浮细胞)

a) 细胞准备:按照标准方法培养细胞,必须保证检测用细胞状态。按照适当方法,清洗并收集足量的细胞。

b) 药物诱导:将收集好的细胞悬浮于适量稀释好的药物,于37℃细胞培养箱内避光孵育,实际诱导时间由药物特性和细胞类型决定。

c) (可选)阳性对照:先用无血清培养基稀释阳性对照(Rosup, 100 mM)到常用工作浓度100 μM,加入细胞,37℃避光孵育0.5~4 h 以提高活性氧水平,不同细胞类型存在差异。例如:HeLa 细胞需孵育30-60 min,MRC5 人胚胎成纤维细胞则需孵育90min。

d) ROS 探针准备:探针装载前,按照1:1000 用无血清培养液稀释DCFH-DA,使其终浓度为10 μM。

e) 探针装载:除去细胞内药物,离心收集细胞,加入稀释好的探针,使其细胞密度为1×10^6 ~ 2×10^7。

注意:细胞密度需根据后续的检测体系,检测方法,以及检测总量来进行调整。例如:对于流式分析,单管检测内细胞数目不少于10^4,也不可多于10^6。

f) 细胞清洗:用无血清细胞培养液洗涤细胞1-2 次,以充分去除未进入细胞内的DCFH-DA。

2. 荧光显微照相操作方法

a) 对贴壁生长细胞或活组织,可直接在荧光显微镜下观察;对悬浮生长细胞,取25-50 μL 细胞悬液滴到一张显微载玻片上,再盖上一张盖玻片。

b) 荧光显微镜下,选用FITC 滤光片观察荧光,去除背景观察荧光的变化。

3. 流式细胞分析操作方法

a) 对贴壁生长细胞,用胰酶消化制备成单细胞悬液;对悬浮生长细胞,直接收集细胞。用0.5-1 mL PBS 重悬细胞(0.5~1 x 10^5/ml)。

b) 选择流式细胞仪FL1 或BL1 通道,488nm 激发,测定530nm 的发射,细胞应可分成两个亚群:ROS 阴性细胞仅有很低的荧光强度,ROS 阳性细胞有较强的绿色荧光。

4.参数设置

使用488nm激发波长,525nm发射波长,实时或逐时间点检测刺激前后荧光的强弱。DCF的荧光光谱和FITC非常相似,可以用FITC的参数设置检测DCF。DCF的激发光谱和发射光谱参考下图。
ROS 活性氧检测试剂盒

使用活性氧检测试剂盒(Reactive oxygen species assay kit)显示CHO细胞内活性氧荧光。

左图:CHO细胞用试剂盒配备的活性氧阳性对照处理;右图:正常CHO细胞。绿色荧光表明细胞活性氧急剧增加,并能显示其定位。

Product composition

 

Product name

Package

Liquid A

H2DCFH-DA(10mM)

0.1mL

Liquid B

Active oxygen positive control(Rosup,50mg/mL)

1mL

 

Product Introduction

Reactive Oxygen Species Assay Kit (Reactive Oxygen Species Assay Kit) is a kit that uses fluorescent probe H2DCFH-DA for reactive oxygen detection. H2DCFH-DA itself has no fluorescence and can freely pass through the cell membrane. After entering the cell, it can be hydrolyzed by intracellular esterase to produce DCFH. DCFH cannot penetrate the cell membrane, so that the probe is easily loaded into the cell. The reactive oxygen species in cells can oxidize non-fluorescent DCFH to produce fluorescent DCF. Detecting the fluorescence of DCF can know the level of reactive oxygen species in the cell. According to the fluorescence produced in living cells, the content and changes of reactive oxygen species can be judged. It can be directly observed with a flow cytometer or a fluorescence microscope. It is a classic method for detecting reactive oxygen species in tissues or living cells. This kit provides Rosup, a positive control reagent for active oxygen, to facilitate the detection of active oxygen. Rosup is a mixture with a concentration of 50 mg/mL. Rosup is a reactive oxygen-inducing drug. According to its fluorescence signal intensity, the true level of reactive oxygen can be analyzed.

The kit has low background, high sensitivity, wide linear range and convenient use. This kit can measure 1000 samples 1000T (96 well plate).

Precautions

1. After the probe is loaded, be sure to wash the remaining probes that have not entered the cell, otherwise it will cause a high background.

2. The positive control Rosup generally uses a concentration of 100 μM (the recommended concentration is 100-400 μM, depending on the cell type). Usually, a significant increase in reactive oxygen species can be observed 0.5-4h after stimulation. For different cells, the effect of reactive oxygen species positive control may be quite different. If no increase in active oxygen is observed within 30 minutes after stimulation, the induction time can be extended or the concentration of active oxygen positive control can be appropriately increased. If the reactive oxygen species increase too quickly, shorten the induction time or appropriately reduce the concentration of the reactive oxygen species positive control.

3. For some special cells, if the negative control cells without stimulation are found to have stronger fluorescence during the experiment, you can dilute DCFH-DA at 1:2000-1:5000 to make the final concentration of DCFH-DA 2-5 μM. The loading time of the probe can also be appropriately adjusted within 15-60 min according to the situation.

4. Active oxygen positive control (Rosup) is only used as a positive control sample, and it is not necessary to add active oxygen positive control to every sample.

5. After the probe is loaded and the remaining probes are cleaned, scan the excitation wavelength and emission wavelength to confirm whether the probe is well loaded. Please refer to the figure above for the excitation and emission spectra of DCF.

6. Try to shorten the time from probe loading to measurement (except stimulation time) to reduce all possible errors.

7. For your safety and health, please wear lab coats and disposable gloves.

8. If it is quantitative, make a standard curve. First make a different concentration of H2O2 oxidized DCFA fluorescence value, make a standard curve, the X axis is the H2O2 concentration, the y axis is the fluorescence value, and get an equation. Look at the fluorescence value of your sample, that is, the Y value, and the corresponding X value Yes.

9. After some cells are loaded with probes, the cells are easy to float up, and the experimental group will suck some of the cells when washing the cells. Therefore, the cell mass is doubled when the cells are planted, so that the cells are tightly connected and adhere to the wall more firmly, and the fluorescence value of the experimental group is higher. In addition, H2DCFDA is very sensitive. The concentration of the working solution should be lower, 1-2μM is enough. If the concentration is too high, non-specific staining may occur. This probe is very unstable. Once it is oxidized, the background fluorescence value will increase. It is best to use the working solution and prepare it.

Instructions for use

1. Load ROS probe

1.1 In-situ loading of probes (only for adherent cells)

a) Cell preparation: Plating cells the day before the test to ensure that the number of cells during the test is less than 5×10^5/ml.

b) Drug induction: Remove the cell culture medium, add the drug treatment diluted with serum-free medium, and incubate in a 37°C cell culture incubator in the dark. The actual induction time is determined by the characteristics of the drug and the cell type.

c) (Optional) Positive control: First dilute the positive control (Rosup, 100 mM) with serum-free medium, etc. to a common working concentration of 100 μM, add cells, and incubate at 37°C for 0.5 to 4 h in the dark to increase the level of reactive oxygen species , Different cell types are different. For example: HeLa cells need to be incubated for 30-60 minutes, and MRC5 human embryonic fibroblasts need to be incubated for 90 minutes.

d) ROS probe preparation: Before loading the probe, dilute DCFH-DA with serum-free culture medium at a ratio of 1:1000 to make the final concentration 10 μM.

e) ROS probe loading: aspirate the treatment drug, and add DCFH-DA working solution diluted in an appropriate volume. The added volume must cover the cells sufficiently. For example, a 6-well plate is usually no less than 1000 μL, and a 96-well plate is usually no less than 100 μL. Incubate for 30 min in a cell culture box at 37°C in the dark.

f) Cell washing: Wash the cells with serum-free culture medium 1 to 2 times to fully remove the DCFH-DA that has not entered the cells.

1.2 Load probes after collecting cells (applicable to adherent cells and suspension cells)

a) Cell preparation: Cells are cultured according to standard methods, and the cell status for testing must be guaranteed. According to the appropriate method, wash and collect enough cells.

b) Drug induction: Suspend the collected cells in an appropriate amount of diluted drug, and incubate in a 37°C cell incubator in the dark. The actual induction time is determined by the characteristics of the drug and the cell type.

c) (Optional) Positive control: First dilute the positive control (Rosup, 100 mM) with serum-free medium to a common working concentration of 100 μM, add cells, and incubate at 37°C in the dark for 0.5 to 4 hours to increase the level of reactive oxygen species. There are differences in cell types. For example: HeLa cells need to be incubated for 30-60 minutes, and MRC5 human embryonic fibroblasts need to be incubated for 90 minutes.

d) ROS probe preparation: Before loading the probe, dilute DCFH-DA with serum-free culture medium at a ratio of 1:1000 to make the final concentration 10 μM.

e) Probe loading: remove the intracellular drugs, collect the cells by centrifugation, and add the diluted probe to make the cell density 1×10^6~2×10^7.

Note: The cell density needs to be adjusted according to the subsequent detection system, detection method, and total detection volume. For example, for flow cytometry, the number of cells in a single tube is not less than 10^4 and not more than 10^6.

f) Cell washing: Wash the cells with serum-free cell culture medium 1-2 times to fully remove the DCFH-DA that has not entered the cells.

2. Fluorescence microscopy operation method

a) For adherent growth cells or living tissues, you can directly observe under a fluorescence microscope; for suspended growth cells, drop 25-50 μL of cell suspension onto a microscope slide, and then cover with a cover glass.

b) Under a fluorescence microscope, use FITC filters to observe fluorescence, and remove background to observe changes in fluorescence.

3. Flow cytometry operation method

a) For adherent growth cells, trypsinization is used to prepare a single cell suspension; for suspension growth cells, the cells are directly collected. Resuspend the cells with 0.5-1 mL PBS (0.5-1 x 10^5/ml).

b) Choose the FL1 or BL1 channel of the flow cytometer, excite at 488nm, and measure the emission at 530nm. Cells should be divided into two subgroups: ROS negative cells have only very low fluorescence intensity, and ROS positive cells have strong green fluorescence.

4. Parameter setting

Using 488nm excitation wavelength, 525nm emission wavelength, real-time or time-by-time detection of fluorescence intensity before and after stimulation. The fluorescence spectrum of DCF is very similar to FITC, and the parameters of FITC can be used to detect DCF. Refer to the figure below for the excitation and emission spectra of DCF.

ROS 活性氧检测试剂盒

Reactive oxygen species assay kit was used to display the fluorescence of reactive oxygen species in CHO cells.

Left image: CHO cells were treated with the reactive oxygen species included in the kit; right image: normal CHO cells. Green fluorescence indicates a sharp increase in cellular reactive oxygen species and can show its location.

相关属性

储存温度 避光,-20°C储存
品牌 Jinpan

Qubit dsDNA HS 定量检测试剂盒

Qubit dsDNA HS 定量检测试剂盒

有货

Qubit dsDNA HS 定量检测试剂盒

品牌:Jinpan
Qubit dsDNA HS Assay Kit

MSDS

质检证书(CoA)

相似产品

货号 (SKU) 包装规格 是否现货 价格 数量
Q266241-2000T 2000T 期货 Qubit dsDNA HS 定量检测试剂盒  
Q266241-200T×10 200T×10 期货 Qubit dsDNA HS 定量检测试剂盒  

基本信息

产品名称 Qubit dsDNA HS 定量检测试剂盒
英文名称 Qubit dsDNA HS Assay Kit
运输条件 冰袋运输

一般描述

产品介绍

Qubit dsDNA HS分析试剂盒(Qubit dsDNA HS Assay Kit) 专门设计用于在Qubit 荧光仪, 但也可以在任何荧光计或荧光酶标仪中使用。该试剂盒对双链DNA(dsDNA)有高度的选择性,不会定量RNA,可以定量浓度为10 pg/µl 到100 ng/µl的样品。试剂盒提供了浓缩的分析试剂,稀释缓冲液和预制的DNA标准。您只需用提供的缓冲液稀释试剂,添加样品(体积在 1 µL 到 20 µL 范围内), 然后使用Qubit荧光仪读取浓度。该试剂盒对于常见污染物,如盐,游离的核苷酸,溶剂,去污剂,蛋白质的耐受性极好。

HS (High Sensitivity):高灵敏度

搭配仪器

对于1-20个样品:Qubit dsDNA HS定量试剂盒和Qubit荧光仪一起使用

对于20-2000个样品:Quant-iT dsDNA HS定量试剂盒和酶标仪(microplate reader)一起使用

注意事项

1.所有Qubit定量试剂盒都可以与Qubit 1.0,Qubit 2.0,Qubit 3和Qubit 4荧光计一起使用。

2.需要500μL薄壁PCR管,但不包括在内。

3.也可作为1X dsDNA定量试剂盒,配好的试剂可作为即用型溶液稳定长达一年。

产品特点

核心范围(高可信度):1 ng / mL至500 ng / mL

扩展范围(中等可信度):0.5 ng / mL至1 ng / mL和500 ng / mL至600 ng / mL

用于定量PCR产物、病毒DNA和亚克隆(subcloning)样品

RNA,盐,溶剂,蛋白质和游离核苷酸的存在下是准确的

产品参数

Ex(nm) 488        Em(nm) 520

自备材料

用于混合Qubit工作溶液的塑料容器(一次性)

Qubit  Assay Tubes分析试管(500管)或Axygen PCR-05-C管(VWR)

Product description

Qubit dsDNA HS Assay Kit (Qubit dsDNA HS Assay Kit) is specially designed to be used in Qubit Fluorometer, but it can also be used in any fluorometer or fluorescence microplate reader. The kit is highly selective for double-stranded DNA (dsDNA), does not quantify RNA, and can quantify samples with concentrations ranging from 10 pg/µl to 100 ng/µl. The kit provides concentrated analysis reagents, dilution buffer and pre-made DNA standards. All you need to do is to dilute the reagent with the provided buffer, add the sample (volume in the range of 1 µL to 20 µL), and then use the Qubit Fluorometer to read the concentration. The kit has excellent tolerance to common contaminants such as salts, free nucleotides, solvents, detergents, and proteins.

HS (High Sensitivity): High sensitivity

Matching instrument

For 1-20 samples: Qubit dsDNA HS Quantitative Kit and Qubit Fluorometer are used together

For 20-2000 samples: Quant-iT dsDNA HS quantitative kit and microplate reader are used together

Precautions

1. All Qubit quantitative kits can be used with Qubit 1.0, Qubit 2.0, Qubit 3 and Qubit 4 fluorometers.

2. A 500μL thin-walled PCR tube is required, but not included.

3. It can also be used as a 1X dsDNA quantitative kit. The prepared reagents can be used as a ready-to-use solution that is stable for up to one year.

Features

Core range (high confidence): 1 ng / mL to 500 ng / mL

Extended range (medium confidence): 0.5 ng/mL to 1 ng/mL and 500 ng/mL to 600 ng/mL

Used for quantitative PCR products, viral DNA and subcloning samples

It is accurate in the presence of RNA, salt, solvent, protein and free nucleotide

Product parameter

Ex(nm) 488    Em(nm) 520

Bring your own materials

Plastic container for mixing Qubit working solution (disposable)

Qubit Assay Tubes (500 tubes) or Axygen PCR-05-C tubes (VWR)

相关属性

储存温度 2-8°C储存,避光,干燥
品牌 Jinpan

PicoGreen dsDNA 定量检测试剂盒

PicoGreen dsDNA 定量检测试剂盒

有货

PicoGreen dsDNA 定量检测试剂盒

品牌:Jinpan
PicoGreen dsDNA Assay Kit

MSDS

质检证书(CoA)

相似产品

货号 (SKU) 包装规格 是否现货 价格 数量
P266245-200T×10 200T×10 期货 PicoGreen dsDNA 定量检测试剂盒  
P266245-2000T 2000T 期货 PicoGreen dsDNA 定量检测试剂盒  

基本信息

产品名称 PicoGreen dsDNA 定量检测试剂盒
英文名称 PicoGreen dsDNA Assay Kit
运输条件 冰袋运输

一般描述

产品介绍

用于 dsDNA 定量检测的 PicoGreen荧光染料

1.应用说明

在分子生物学的试验过程中,PicoGreen dsDNA 定量试剂盒是荧光检测双链 DNA 并进行定量 的一种产品,这种方法非常灵敏。常用于分子生物学技术中的:cDNA 文库的构建;用于亚克隆的  DNA 片段纯化及应用,比如进行 DNA 定量、产物扩增和引物的进一步检测。 疫苗是现代疾病预防中常用的控制方式。如今许多疫苗是细胞培养疫苗,比如重组乙肝疫苗、狂犬病疫苗等大多数疫苗都采用细胞培养的方法生产。其中,疫苗的纯化是关键问题,我们需要尽可能的去除宿主细胞DNA和宿主蛋白。假若宿主细胞的DNA和蛋白同疫苗一起注入人体将会产生不可预料的后果。

常规的DNA含量的检测方法是在260nm(A260)处测其吸光值。这种方法的主要缺点是核苷酸、单链核酸和蛋白质对信号的影响很大,并且还会受到核酸制备过程中污染物的干扰,无法区分DNA和RNA,而且这种方法不灵敏(5μg/mLdsDNA溶液A260=0.1)。PicoGreen定量检测方法简单、方便,被多家生物制品厂所选择,成为生物制品残留DNA检测的标准。

目前该方法已纳入2010年版《中国药典》

原理:

PicoGreen与DNA双链结合后才发出的荧光,无DNA不发荧光;所发荧光与DNA浓度成正比。在2010年《中国药典》中提出,PicoGreen定量DNA的方法检出限约0.3ng/ml,DNA含量在1.25-80ng/mL范围时线性较好(R2>0.99).

优点:

1)该方法可以测定来源于任何表达宿主样品中的双链DNA。

2)可以直接定量PCR扩增产物而无需从反应混合物中纯化DNA。

3)远远超出传统紫外A260的检测方法和Hoechst33258的灵敏度。

4)较高浓度的盐,尿素,乙醇,氯仿,去垢剂,蛋白或琼脂糖对测定无影响。

5)在等摩尔浓度 ssDNA 和 RNA 存在的条件下测定 dsDNA,其影响很小。

2.所需器材

• 微型荧光计;便携式荧光仪-上海互帼科学仪器有限公司HG-9型;1cm石英比色皿

• PicoGreen dsDNA 定量检测试剂盒, 1mL 单位量的试剂浓缩液足够 2mL 体积的 200 次测定。

1×TE(10mM Tris 1mM EDTA)pH8.0; 250ug/mL Sigma 小牛胸腺DNA

3.实验方案

试剂制备

PicoGreen dsDNA 定量试剂是以 1mL 的浓缩液形式保存在无水的 DMSO(二甲基亚砜)中。实 验当天,配制 2XPicoGreen 试剂的操作溶液,用 1xTE 按 1:200 的比例稀释浓缩液(10mM Tris-HCl,1mM EDTA,pH7.5)。如果要准备足够的操作溶液测定 20 个样品,可在 20mL1x TE 中加入 100μL PicoGreen dsDNA 定量试剂。由于试剂容易吸附到玻璃表面,要在塑料容器中配制。PicoGreen 试剂 见光易降解,所以应将配好的溶液用箔包住或放置暗处避光保存。

溶液最好在配制好数小时内使用,以保证最佳结果。

实验方法:

1). 标准品工作液的配制:

 Sigama小牛胸腺嘧啶DNA干粉(货号:D4522-1MG)1mg(Tris,Nacl等浓度已成标准体系),加入1mL双蒸水,配制成1mg∕mL的标准品工作液;

2). 染料工作液的配置:

6 uLPicoGreen加入1mL TE(注意:用1×TE将PicoGreen稀释200倍,现用现配,注意避光)。

3). 标准品工作液稀释:

1)母液稀释:取10ul(1mg∕mL)标准品工作液加入到990ul TE溶液中,浓度稀释成10ug∕mL,取10ul(10ug∕mL)标准品工作液加入到990ul TE溶液中,浓度稀释成100ng∕mL;

2)倍比稀释:取800ul (100ng∕mL)的标准品工作液加入到200ul TE溶液中,浓度达到80ng∕mL(药典规定:荧光染色方法DNA含量在1.25-80 ng/mL范围线性较好, 该法DNA检出限为0.3 ng/ml),取500ul(80ng∕mL)的标准品工作液加入到500ul TE溶液中,浓度稀释到40ng∕mL;依次倍比稀释,配成20ng/ml、10ng/ml、5.0ng/ml、2.5ng/ml、1.25ng/ml、0.625ng/ml的标准品溶液;

4).标准曲线的制备:倍比稀释后的各梯度标准品溶液和染料工作液各取100ul混匀,避光室温放置5min。使用FB-15型便携式荧光仪检测样品的荧光值:将混合后的溶液加入微量比色皿,确信不要在样品中引入气泡,轻轻地弹微量检测皿的外部,可以驱散气泡。以1×TE缓冲液为blank,测定样品和空白对照的荧光值;用标准品溶液的浓度(ng/ml)对应的荧光强度作直线回归,制备标准曲线。

5). 测量剩余样品的荧光值。荧光计将给出一个直接的浓度读数,数据可以用来产生DNA 浓度的标准曲线。

PicoGreen dsDNA 定量检测试剂盒


产品参数

Ex(nm) 488      Em(nm) 520

Product description

PicoGreen fluorescent dye for dsDNA quantitative detection

1. Application description

In molecular biology experiments, the PicoGreen dsDNA quantification kit is a product for fluorescent detection and quantification of double-stranded DNA. This method is very sensitive. Commonly used in molecular biology technology: cDNA library construction; DNA fragment purification and application for subcloning, such as DNA quantification, product amplification, and further primer detection. Vaccines are a commonly used control method in modern disease prevention. Many vaccines today are cell culture vaccines, such as recombinant hepatitis B vaccine, rabies vaccine and most of the vaccines are produced using cell culture methods. Among them, the purification of vaccines is a key issue, and we need to remove host cell DNA and host proteins as much as possible. If the host cell’s DNA and protein are injected into the human body together with the vaccine, it will have unpredictable consequences.

The conventional DNA content detection method is to measure its absorbance at 260nm (A260). The main disadvantage of this method is that nucleotides, single-stranded nucleic acids and proteins have a great influence on the signal, and are also interfered by contaminants in the nucleic acid preparation process, and cannot distinguish between DNA and RNA, and this method is not sensitive (5μg) /mLdsDNA solution A260=0.1). The PicoGreen quantitative detection method is simple and convenient, and has been selected by many biological product manufacturers, and has become the standard for the detection of residual DNA in biological products. At present, this method has been included in the 2010 edition of “Chinese Pharmacopoeia”

Principle:

The fluorescence emitted after PicoGreen is combined with the double strand of DNA, without DNA, does not emit fluorescence; the fluorescence emitted is proportional to the concentration of DNA. It was proposed in the Chinese Pharmacopoeia in 2010 that the detection limit of PicoGreen method for quantifying DNA is about 0.3ng/ml, and the linearity is better when the DNA content is in the range of 1.25-80ng/mL (R2>0.99).

Advantage:

1) This method can measure double-stranded DNA in samples derived from any expression host.

2) It is possible to directly quantify PCR products without purifying DNA from the reaction mixture.

3) Far beyond the traditional UV A260 detection method and the sensitivity of Hoechst33258.

4) Higher concentrations of salt, urea, ethanol, chloroform, detergent, protein or agarose have no effect on the determination.

5) Measuring dsDNA in the presence of equimolar concentrations of ssDNA and RNA has little effect.

2. Required equipment

• Mini Fluorometer; Portable Fluorometer-Shanghai Hubei Scientific Instrument Co., Ltd. HG-9; 1cm quartz cuvette

• PicoGreen dsDNA Quantitative Detection Kit, a 1mL unit volume of reagent concentrate is sufficient for 200 determinations in a volume of 2mL.

1×TE (10mM Tris 1mM EDTA) pH8.0; 250ug/mL Sigma calf thymus DNA

3. Experimental protocol

Reagent preparation

PicoGreen dsDNA quantification reagent is stored in anhydrous DMSO (dimethyl sulfoxide) in the form of a 1mL concentrated solution. On the day of the experiment, prepare the operating solution of 2XPicoGreen reagent and dilute the concentrated solution (10mM Tris-HCl, 1mM EDTA, pH7.5) with 1xTE at a ratio of 1:200. If you want to prepare enough operating solution to measure 20 samples, you can add 100μL PicoGreen dsDNA quantification reagent to 20mL1x TE. Since the reagent is easily adsorbed to the glass surface, it should be prepared in a plastic container. PicoGreen reagent is easily degraded when exposed to light, so the prepared solution should be wrapped in foil or stored in a dark place away from light.

The solution is best used within a few hours of preparation to ensure the best results.

Experimental method:

1). Preparation of standard working solution:

Sigama calf thymine DNA dry powder (article number: D4522-1MG) 1mg (Tris, Nacl and other concentrations have become standard systems), add 1mL of double distilled water to prepare 1mg/mL standard working solution;

2). Dye working solution configuration:

6 Add 1mL TE to uLPicoGreen (Note: Dilute PicoGreen 200 times with 1×TE, and use it now, please avoid light).

3). Standard working solution dilution:

(1) Mother liquor dilution: Take 10ul (1mg/mL) standard working solution and add it to 990ul TE solution, dilute to 10ug/mL, take 10ul (10ug/mL) standard working solution and add it to 990ul TE solution, concentration Dilute to 100ng∕mL;

(2) Multiple dilution: Take 800ul (100ng/mL) standard working solution and add it to 200ul TE solution, the concentration reaches 80ng/mL (pharmacopoeia stipulates: the DNA content of fluorescent staining method is linear in the range of 1.25-80 ng/mL Ok, the DNA detection limit of this method is 0.3 ng/ml). Take 500ul (80ng/mL) of standard working solution and add it to 500ul TE solution, and dilute to 40ng/mL; successively dilute by multiples and make 20ng/mL. Standard solution of ml, 10ng/ml, 5.0ng/ml, 2.5ng/ml, 1.25ng/ml, 0.625ng/ml;

4). Preparation of standard curve: take 100ul of each gradient standard solution and dye working solution diluted by multiple ratios and mix them evenly, and place them at room temperature in the dark for 5 minutes. Use the FB-15 portable fluorometer to detect the fluorescence value of the sample: add the mixed solution into the micro cuvette, make sure not to introduce air bubbles into the sample, and gently flick the outside of the micro detection cuvette to disperse the air bubbles. Use 1×TE buffer as blank to measure the fluorescence values ​​of the sample and the blank control; use the fluorescence intensity corresponding to the concentration of the standard solution (ng/ml) to make a linear regression to prepare a standard curve.

5). Measure the fluorescence value of the remaining samples. The fluorometer will give a direct concentration reading, and the data can be used to generate a standard curve of DNA concentration.
PicoGreen dsDNA 定量检测试剂盒


Product parameter

Ex(nm) 488        Em(nm) 520

相关属性

储存温度 2-8°C储存,避光,干燥
品牌 Jinpan

OligoGreen ssDNA 定量检测试剂盒

OligoGreen ssDNA 定量检测试剂盒

有货

OligoGreen ssDNA 定量检测试剂盒

品牌:Jinpan
OligoGreen ssDNA quantitative detection kit

MSDS

质检证书(CoA)

相似产品

货号 (SKU) 包装规格 是否现货 价格 数量
O273080-2000T 2000T 期货 OligoGreen ssDNA 定量检测试剂盒  
O273080-200T×10 200T×10 期货 OligoGreen ssDNA 定量检测试剂盒  

基本信息

产品名称 OligoGreen ssDNA 定量检测试剂盒
英文名称 OligoGreen ssDNA quantitative detection kit
运输条件 冰袋运输

一般描述

OliGreen寡核苷酸定量试剂是一种超灵敏的荧光核酸染料,用于定量溶液中的寡核苷酸和单链DNA(ssDNA。使用OliGreen ssDNA定量试剂从0.1到1000 ng / mL的合成24-mer(M13测序引物)线性定量。在480 nm下激发10 mm°C 10 mm比色皿中的样品。使用分光荧光计在520 nm处测量荧光发射强度,并绘制为寡核苷酸浓度的函数。寡核苷酸浓度在0到2.0 ng / mL之间获得的结果的扩大。

OliGreen Oligonucleotide Quantitative Reagent is an ultra-sensitive fluorescent nucleic acid dye that is used to quantify oligonucleotides and single-stranded DNA (ssDNA) in solution. Use OliGreen ssDNA Quantitative Reagent for synthesis from 0.1 to 1000 ng/mL 24- mer (M13 sequencing primer) linear quantification. Excite the sample in a 10 mm°C 10 mm cuvette at 480 nm. Measure the fluorescence emission intensity at 520 nm using a spectrofluorometer and plot it as a function of oligonucleotide concentration The expansion of the results obtained with oligonucleotide concentrations between 0 and 2.0 ng/mL.

相关属性

储存温度 2-8°C储存,避光,干燥
品牌 Jinpan

Calcein AM /PI试剂盒

Calcein AM /PI试剂盒

有货

Calcein AM /PI试剂盒

品牌:Jinpan
Calcein AM /PI Kit

MSDS

质检证书(CoA)

相似产品

货号 (SKU) 包装规格 是否现货 价格 数量
C272922-500T 500T 期货 Calcein AM /PI试剂盒  

基本信息

产品名称 Calcein AM /PI试剂盒
英文名称 Calcein AM /PI Kit
运输条件 冰袋运输

一般描述

产品介绍

钙黄绿素-AM (Calcein-AM) 和碘化丙啶 (PI) 溶液,分别对活细胞和死细胞染色,可用于同时对活细胞和死细胞进行荧光染色。Calcein-AM的乙酸甲基酯亲脂性很高,使其可透过细胞膜。尽管Calcein-AM本身并不是荧光分子,但通过活细胞内的酯酶作用,Calcein-AM能脱去AM基,产生的Calcein能发出强绿色荧光 (激发: 490 nm,发射: 515 nm)。因此Calcein-AM仅对活细胞染色。另一方面,作为核染色染料的PI不能穿过活细胞的细胞膜。它穿过死细胞膜的无序区域而到达细胞核,并嵌入细胞的DNA双螺旋从而产生红色荧光 (激发: 535 nm,发射: 617 nm)。由于Calcein和PI-DNA都可被490 nm激发,因此可用荧光显微镜同时观察活细胞和死细胞。用545 nm激发,仅可观察到死细胞。由于不同细胞系的最佳染色条件不同,我们建议个别确定Calcein-AM和PI的合适浓度。

I. 试剂  Calcein-AM 2mM 50uL in DMSO;  PI (1.5mM) 150uL in water

II. 用荧光显微镜观察细胞形态

HeLa细胞染色为例,请注意不同的细胞种类、不同浓度,有不同的观察条件。根据细胞条件,摸索不同条件下的细胞贴壁情况和试剂浓度的配制等最佳条件。

1. 染色溶液的配制

1) 用1 ml无水DMSO溶解2 mg Calcein-AM,制备成2 mmol/l的Calcein-AM储备液,-20℃下密闭冷冻保存。

2) 用1 ml ddH2O溶解2 mg PI,制备成3 mmol/l的PI储备液,-20度下密闭冷冻保存,可以保存一年。

3) 将Calcein-AM储备液和PI储备液放置于室温。

4) 加5 µl Calcein-AM储备液和15 µl PI储备液至5 ml PBS中配制成染色溶液。Calcein-AM的终浓度为2 µmol/l,PI的终浓度为4.5 µmol/l。

2. 细胞染色

1) 染色HeLa细胞等贴壁细胞时,先用Trypsin-EDTA等消化细胞,制备成细胞悬液。

2) 将细胞悬液离心3分钟 (1,000 rpm)。

3) 去除上清液,加入PBS缓冲液,细胞数量调整至10e5-10e6个/ml。再用移液器充分混匀。

4) 由于培养基中的血清等含有酯酶,Calcein-AM遇水会分解,会导致空白上升,所以需要离心数次,用PBS洗涤数次直到完全洗净。

5) 将200 µl细胞悬液移至小试管中,加入100 µl染色溶液,在37℃下孵育15分钟。

6) 在盖玻片上滴加适量的染色的细胞溶液。

7) 在荧光显微镜下,先用490±10 nm波长激发,观察黄绿色的活细胞,还可以同时观察到红色的死细胞,然后用545 nm波长激发,能够看到红色的死细胞。

3. 染色试剂的最佳浓度

Calcein-AM和PI最佳浓度根据不同的细胞种类而定,通过以下的操作,我们可以找到不同细胞染色试剂的最佳浓度。

1) 通过在0.1%皂苷或0.1-0.5%毛地黄皂苷中孵育10分钟或通过在70%乙醇中孵育30分钟制备死细胞。

2) 用0.1-10 µM PI溶液对死细胞染色,以便找到仅对细胞核染色而不对细胞质染色的PI浓度。

3) 用0.1-10 µM Calcein-AM溶液对死细胞染色,以便找到不对细胞质染色的Calcein-AM浓度。接着用该浓度的Calcein-AM对活细胞染色以检验活细胞可否被染色。

III. 注意事项

1) Calcein-AM的ester部位遇到湿气会分解,使用后请在-20度下密闭冷冻保存,防止水分进入。Calcein-AM储备液用缓冲液或培养基等稀释时尽量现配现用。

2) 使用时一定要带手套、眼罩、口罩。万一接触到皮肤的话,迅速使用大量水清洗。

Product description

Calcein-AM (Calcein-AM) and propidium iodide (PI) solutions stain live and dead cells respectively, and can be used to stain live and dead cells simultaneously. The methyl acetate of Calcein-AM is highly lipophilic, allowing it to penetrate cell membranes. Although Calcein-AM itself is not a fluorescent molecule, through the action of esterase in living cells, Calcein-AM can remove the AM group, and the generated Calcein can emit strong green fluorescence (excitation: 490 nm, emission: 515 nm). Therefore, Calcein-AM only stains live cells. On the other hand, PI, which is a nuclear staining dye, cannot pass through the cell membrane of living cells. It passes through the disordered area of ​​the dead cell membrane to reach the cell nucleus, and is embedded in the cell’s DNA double helix to generate red fluorescence (excitation: 535 nm, emission: 617 nm). Since both Calcein and PI-DNA can be excited at 490 nm, a fluorescence microscope can be used to observe live and dead cells simultaneously. With 545 nm excitation, only dead cells can be observed. Due to the different optimal staining conditions for different cell lines, we recommend determining the appropriate concentration of Calcein-AM and PI individually.

I. Reagent   Calcein-AM 2mM 50uL in DMSO;  PI (1.5mM) 150uL in water

II. Observe cell morphology with a fluorescence microscope

Take HeLa cell staining as an example. Please note that different cell types and different concentrations have different observation conditions. According to the cell conditions, explore the best conditions for cell adhesion and reagent concentration under different conditions.

1. Preparation of dyeing solution

1) Dissolve 2 mg of Calcein-AM with 1 ml of anhydrous DMSO to prepare a 2 mmol/l Calcein-AM stock solution, and store it in a tightly sealed manner at -20°C.

2) Dissolve 2 mg of PI with 1 ml of ddH2O to prepare a 3 mmol/l PI stock solution. Store it in a tightly sealed manner at -20°C, which can be stored for one year.

3) Put the Calcein-AM stock solution and PI stock solution at room temperature.

4) Add 5 µl Calcein-AM stock solution and 15 µl PI stock solution to 5 ml PBS to prepare a staining solution. The final concentration of Calcein-AM is 2 µmol/l and the final concentration of PI is 4.5 µmol/l.

2. Cell staining

1) When staining adherent cells such as HeLa cells, first digest the cells with Trypsin-EDTA, etc. to prepare a cell suspension.

2) Centrifuge the cell suspension for 3 minutes (1,000 rpm).

3) Remove the supernatant, add PBS buffer, and adjust the number of cells to 10e5-10e6 cells/ml. Mix thoroughly with a pipette.

4) Since the serum in the culture medium contains esterase, Calcein-AM will decompose when exposed to water, which will cause the blank to rise, so it needs to be centrifuged several times and washed several times with PBS until it is completely washed.

5) Transfer 200 µl of cell suspension to a small test tube, add 100 µl of staining solution, and incubate at 37°C for 15 minutes.

6) Drop an appropriate amount of stained cell solution on the cover glass.

7) Under a fluorescence microscope, first excite with a wavelength of 490±10 nm to observe the yellow-green live cells, and also observe the red dead cells at the same time, and then excite with the 545 nm wavelength to see the red dead cells.

3. Optimal concentration of staining reagent

The optimal concentration of Calcein-AM and PI depends on different cell types. Through the following operations, we can find the optimal concentration of different cell staining reagents.

1) Prepare dead cells by incubating in 0.1% saponin or 0.1-0.5% digitonin for 10 minutes or by incubating in 70% ethanol for 30 minutes.

2) Stain dead cells with 0.1-10 µM PI solution to find the PI concentration that only stains the nucleus and not the cytoplasm.

3) Stain dead cells with 0.1-10 µM Calcein-AM solution to find the concentration of Calcein-AM that does not stain the cytoplasm. Then, live cells were stained with this concentration of Calcein-AM to check whether the live cells could be stained.

III. Matters needing attention

1) The ester part of Calcein-AM will decompose when exposed to moisture. After use, please keep it airtight and freeze at -20 degrees to prevent moisture from entering. When the Calcein-AM stock solution is diluted with buffer or culture medium, it should be prepared as soon as possible.

2) Always wear gloves, goggles, and masks when using. In case of contact with the skin, quickly wash with plenty of water.

相关属性

储存温度 2-8°C储存,避光,干燥
品牌 Jinpan

2 × SYBR Green qPCR 试剂盒

2 × SYBR Green qPCR 试剂盒

有货

2 × SYBR Green qPCR 试剂盒

品牌:Jinpan
2 × SYBR Green qPCR Kit

MSDS

质检证书(CoA)

相似产品

货号 (SKU) 包装规格 是否现货 价格 数量
S272945-50T 50T 期货 2 × SYBR Green qPCR 试剂盒  
S272945-200T 200T 期货 2 × SYBR Green qPCR 试剂盒  

基本信息

产品名称 2 × SYBR Green qPCR 试剂盒
英文名称 2 × SYBR Green qPCR Kit
运输条件 超低温冰袋运输

一般描述

产品介绍

组成

50×50μL反应体系

200×50μL反应体系  

2×SYBR Green qPCR Mix

1.25 mL

1.25 mL×4

产品参数

Ex(nm)497

Em(nm)525

储存:-20 ℃ 避光保存6个月,使用前充分溶解混匀。短期使用可放在4 ℃,避免反复冻融。

制品说明
本制品是采用SYBR Green I嵌合荧光法进行Real Time PCR的专用试剂。已经将DNA聚合酶、dNTP、特殊稳定剂、优化的反应缓冲液、BSA和SYBR Green I等试剂预混成一种适合Real Time PCR反应检测用2×Premix Type试剂,具有灵敏度高、特异性强、稳定性好等特点。使用时只需加入模板、引物和水,便可在宽广的定量区域内得到良好的标准曲线,对目的基因进行准确定量检测,重复性好,可信度高。

注意事项

1.本制品不含参比染料ROX,客户可根据qPCR仪器技术指导决定是否需要加ROX参比染料,用于消除信号本底以及校正孔与孔之间产生的荧光信号误差。

2.本制品含SYBR Green I 强光下易分解,降低灵敏度,使用时避免长时间强光照射本制品。

3.建议在冰上配制PCR反应液,再放入PCR仪器中扩增。可以提高扩增特异性,减少背景。

4.本制品含有4 mM MgCl2(反应体系终浓度是2 mM Mg2+),可用25mM MgCl2 优化Mg2+浓度。

建议PCR条件(以50 μL 反应体系为例,反应液配制请在冰上进行)

Components

Volume

Final Concentration

2×SYBR Green qPCR Mix

25μL

DNA Template

2μL

as required

Forward Primer (10μM)

1μL

0.2 μM each

Reverse Primer (10μM)

1μL

0.2 μM each

ddH2O to final volume

50μL

Not applicable

PCR 循环

94℃    2~3 min                     

94℃    10~20 sec

55~65℃  10~20 sec    35~45 cycles

72℃     20~60 sec

72℃    5~10 min  

Product Introduction

Composition

50T×50μL reaction system 

200T×50μL reaction system 

2×SYBR Green qPCR Mix

1.25 mL

1.25 mL×4

Product parameter

Ex(nm)497

Em(nm)525

Storage: Store at -20 ℃ in the dark for 6 months. Dissolve and mix thoroughly before use. It can be stored at 4℃ for short-term use to avoid repeated freezing and thawing.

Product description

This product is a special reagent for Real Time PCR using SYBR Green I chimeric fluorescence method. Reagents such as DNA polymerase, dNTP, special stabilizers, optimized reaction buffer, BSA and SYBR Green I have been premixed into a 2×Premix Type reagent suitable for Real Time PCR reaction detection. It has high sensitivity, strong specificity, Features such as good stability. When using, only need to add template, primer and water, a good standard curve can be obtained in a wide quantitative area, and the target gene can be accurately and quantitatively detected, with good repeatability and high reliability.

Precautions

1. This product does not contain the reference dye ROX. Customers can decide whether to add ROX reference dye according to the qPCR instrument technical guidance to eliminate the signal background and correct the fluorescence signal error generated between the wells.

2. This product contains SYBR Green I. It is easy to decompose under strong light and reduce sensitivity. Avoid long-term strong light exposure to this product.

3. It is recommended to prepare the PCR reaction solution on ice and put it into the PCR instrument for amplification. It can improve amplification specificity and reduce background.

4. This product contains 4 mM MgCl2 (the final concentration of the reaction system is 2 mM Mg2+), and 25mM MgCl2 can be used to optimize the Mg2+ concentration.

Suggested PCR conditions (take a 50 μL reaction system as an example, please prepare the reaction solution on ice)

Components

Volume

Final Concentration

2×SYBR Green qPCR Mix

25μL

DNA Template

2μL

as required

Forward Primer (10μM)

1μL

0.2 μM each

Reverse Primer (10μM)

1μL

0.2 μM each

ddH2O to final volume

50μL

Not applicable

PCR cycle

94℃     2~3 min                     

94℃     10~20 sec

55~65℃   10~20 sec    35~45 cycles

72℃     20~60 sec

72℃    5~10 min  

相关属性

储存温度 避光,-20°C储存
品牌 Jinpan

CUBIC-HV™1 3D 免疫染色试剂盒

CUBIC-HV™1 3D 免疫染色试剂盒

有货

CUBIC-HV™1 3D 免疫染色试剂盒

品牌:Jinpan
CUBIC-HV™1 3D immunostaining kit

MSDS

质检证书(CoA)

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货号 (SKU) 包装规格 是否现货 价格 数量
C406225-1Set 期货 CUBIC-HV™1 3D 免疫染色试剂盒  

基本信息

产品名称 CUBIC-HV™1 3D 免疫染色试剂盒
英文名称 CUBIC-HV™1 3D immunostaining kit
运输条件 冰袋运输

相关属性

敏感性 对热敏感
储存温度 2-8°C储存
品牌 Jinpan

一氧化氮传感器(细胞内)试剂盒(“ NO-ON”)(FL)(可细胞捕获的NO荧光探针)

一氧化氮传感器(细胞内)试剂盒(“ NO-ON”)(FL)(可细胞捕获的NO荧光探针)

有货

一氧化氮传感器(细胞内)试剂盒(“ NO-ON”)(FL)(可细胞捕获的NO荧光探针)

品牌:Jinpan
Nitric Oxide Sensor (Intracellular) Kit (“NO-ON”) (FL) (Cell-trappable NO fluorescent probe)

MSDS

质检证书(CoA)

相似产品

货号 (SKU) 包装规格 是否现货 价格 数量
N356845-1kit 1kit 期货 一氧化氮传感器(细胞内)试剂盒(“ NO-ON”)(FL)(可细胞捕获的NO荧光探针)  

基本信息

产品名称 一氧化氮传感器(细胞内)试剂盒(“ NO-ON”)(FL)(可细胞捕获的NO荧光探针)
英文名称 Nitric Oxide Sensor (Intracellular) Kit (“NO-ON”) (FL) (Cell-trappable NO fluorescent probe)
运输条件 冰袋运输

一般描述

一氧化氮传感器(细胞内)试剂盒(“ NO-ON”)(FL)(可细胞捕获的NO荧光探针)产品详细清单:


货号 产品名 成分货号 成分名称 成分CAS 规格
N356845 一氧化氮传感器(细胞内)试剂盒(“ NO-ON”)(FL)(可细胞捕获的NO荧光探针) C281470-0.5mg 2-{2-氯-6-羟基-5-[2-甲基喹啉-8-基氨甲基]-3-氧代-3H-呫吨-9-基}苯甲酸磺酸 905982-78-7 95%

Nitric Oxide Sensor (Intracellular) Kit (“NO-ON”) (FL) (Cell-trappable NO fluorescent probe) detailed list of products:


Catalog Number Product Name Component Catalog Number Component Name Component CAS Specification&Purity
N356845 Nitric Oxide Sensor (Intracellular) Kit (“NO-ON”) (FL) (Cell-trappable NO fluorescent probe) C281470-0.5mg 2-{2-Chloro-6-hydroxy-5-[2-methylquinolin-8-ylaminomethyl]-3-oxo-3H-xanthen-9-yl}benzoic acid FL 905982-78-7 95%

相关属性

储存温度 2-8°C储存
品牌 Jinpan