| 产品名称 | 1-溴-2,3,4-三苯并yl-L-rhamno吡喃并se |
|---|---|
| 英文名称 | 1-Bromo-2,3,4-tribenzoyl-L-rhamnopyranose |
| 英文别名 | 6-Deoxy-L-mannopyranosyl bromide tribenzoate |
| 运输条件 | 超低温冰袋运输 |
Description
Versatile monosaccharide building block for making rhamnosyl derivatives.
Description
Versatile monosaccharide building block for making rhamnosyl derivatives.
| CAS编号 | 73068-86-7 |
|---|---|
| 储存温度 | -20°C储存 |
| MDL号 | MFCD23704413 |
| 分子量 | 539.37 |
| 品牌 | Jinpan |
| 产品名称 | Cy3 SE |
|---|---|
| 英文名称 | Cy3 SE |
| 运输条件 | 超低温冰袋运输 |
产品简介
菁染料是性能优良的荧光标记染料，摩尔吸光系数在荧光染料中是最高的，其琥珀酰亚胺酯是最常用的脂肪氨基标记试剂，广泛用于蛋白、抗体、核酸及其他生物分子的标记和检测。通过改变次甲基链的长度, 可改变其荧光发射波长，每增加一个双键，按照Huoffman规则正好红移约100nm。
菁染料Cy3和Cy5已成为基因芯片的首选荧光标记物;另外,Cy5, Cy5.5和Cy7的吸收在近红外区背景非常低，是荧光强度最高、最稳定的长波长染料。特别适合于活体小动物体内成像代替放射性元素。
产品参数
Ex(nm) 548 Em(nm) 562 Solvent: Water or DMSO
菁染料琥珀酰亚胺酯（CyDye NHS）的标记
菁染料和生物分子的比例F/P=4～12之间荧光强度最高，F/P值过高荧光探针会自我淬灭并影响生物分子的生物活性，标记生物分子最好是用单琥珀酰亚胺酯，但是用双修饰的CyDye NHS并没有发现交联。CyDye NHS标记抗体在pH (8.5～9.4)时10分钟F/P可达5～6，而在pH 7.0几乎不反应。我们用不同比例的Cy3标记anti-glutathione-S-transferase (GST)多克隆抗体发现用1:1, 5:1, 10:1 和20:1标记时得到的F/P值分别是0.28:1, 1.16:1, 2.3:1 和4.6:1。
操作步骤
水溶性Cy3 NHS标记anti-GST多克隆抗体
市场上买到的抗体如果含有其它蛋白（例如serum albumin或gelatin等）或带氨基的缓冲液会影响标记，在标记前需纯化。
1. 在1 L 0.15 M NaCl 溶液中透析anti-GST抗体(浓度为0.5 mL at 3 mg/mL)，常温透析4小时。
2. 在4°C用新鲜的1 L 0.15 M NaCl 溶液再透析过夜。
3. 第二天用1 L 0.1 M NaHCO3(pH 8.3)透析4小时。
4. 用0.22 μm注射器过滤头过滤抗体溶液。
5. 用0.1 M NaHCO3稀释少量的抗体，在280nm处测其紫外吸收值计算标记抗体的总量（IgG antibody摩尔吸光系数170 000 M-1cm-1at 280 nm).
6. 用DMSO配置Cy3 NHS (MW 765.95)溶液浓度为10 mg/mL；计算所需体积以得到想要的CyDye NHS 和抗体的比值(例如20:1)，然后慢慢将其加入到抗体溶液中，同时在暗处常温缓慢搅拌45分钟。
7. 用1 L 0.15 M NaCl 溶液常温避光透析4小时除去未标记上的Cy3。
8. 在4°C用新鲜的1 L 0.15 M NaCl 溶液再避光透析过夜。
9. 用1 L of 0.01 M PBS/ 0.01% 叠氮化钠溶液常温避光透析4小时, 在4°C常温避光再次透析过夜。
10. 用0.22 μm注射器过滤头过滤抗体溶液。
11. 用0.01 M PBS/0.01 % 叠氮化钠整数倍稀释标记抗体溶液，测量280nm（蛋白）和552nm（Cy）处的紫外可见吸光度。
12. 产品冷冻干燥成粉末或在0.01 M PBS/ 0.01% 叠氮化钠溶液中，-20℃避光储存。
F/P计算：
Cy3在552nm摩尔吸光系数为150000 M-1cm-1；此蛋白在280nm的摩尔吸光系数为170 000 M-1cm-1；不同蛋白的摩尔吸光系数不一样；.Cy3 染料本身在280 nm 的吸收是552nm处的8%。按以下公式计算F/P值。
[Cy3] = A552/150000
[antibody] = {A280- (0.08×A552)}/170000
F/P final= [Cy3]/[antibody]= {1.13×A552}/ {A280- (0.08×A552)}
Product Introduction
Cyanine dye is a fluorescent labeling dye with excellent performance. Its molar absorption coefficient is the highest among fluorescent dyes. Its succinimidyl ester is the most commonly used aliphatic amino group labeling reagent and is widely used for labeling proteins, antibodies, nucleic acids and other biological molecules. And detection. By changing the length of the methine chain, the fluorescence emission wavelength can be changed. For each double bond added, the red shift is exactly 100 nm according to the Huoffman rule.
The cyanine dyes Cy3 and Cy5 have become the first choice fluorescent markers for gene chips; in addition, the absorption of Cy5, Cy5.5 and Cy7 is very low in the near-infrared region, and they are the long-wavelength dyes with the highest fluorescence intensity and the most stable. It is especially suitable for in vivo imaging of small living animals instead of radioactive elements.
Product parameter
Ex(nm) 548 Em(nm) 562 Solvent: Water or DMSO
Labeling of cyanine dye succinimide ester (CyDye NHS)
The ratio of cyanine dye and biomolecule is between F/P=4～12. The fluorescence intensity is the highest. If the F/P value is too high, the fluorescent probe will self-quench and affect the biological activity of biomolecules. It is best to use single amber to label biomolecules. Imide ester, but CyDye NHS modified with double did not find cross-linking. CyDye NHS-labeled antibody can reach 5-6 F/P in 10 minutes at pH (8.5-9.4), but hardly reacts at pH 7.0. We used different ratios of Cy3 to label anti-glutathione-S-transferase (GST) polyclonal antibodies and found that the F/P values obtained when labeled with 1:1, 5:1, 10:1 and 20:1 were 0.28:1 , 1.16:1, 2.3:1 and 4.6:1.
Steps
Water-soluble Cy3 NHS labeled anti-GST polyclonal antibody
Antibodies bought on the market containing other proteins (such as serum albumin or gelatin, etc.) or buffers with amino groups will affect the labeling, and need to be purified before labeling.
1. Dialyze the anti-GST antibody (concentration 0.5 mL at 3 mg/mL) in 1 L 0.15 M NaCl solution for 4 hours at room temperature.
2. Dialysis with fresh 1 L 0.15 M NaCl solution at 4°C overnight.
3. Dialysis with 1 L 0.1 M NaHCO3 (pH 8.3) for 4 hours on the second day.
4. Filter the antibody solution with a 0.22 μm syringe filter.
5. Dilute a small amount of antibody with 0.1 M NaHCO3, measure its UV absorption value at 280nm to calculate the total amount of labeled antibody (IgG antibody molar absorption coefficient 170 000 M-1cm-1at 280 nm).
6. Use DMSO to configure the Cy3 NHS (MW 765.95) solution concentration to 10 mg/mL; calculate the required volume to get the desired ratio of CyDye NHS to antibody (for example, 20:1), and then slowly add it to the antibody solution In the meantime, stir slowly for 45 minutes at room temperature in the dark.
7. Dialysis with 1 L 0.15 M NaCl solution at room temperature and avoid light for 4 hours to remove unlabeled Cy3.
8. Use fresh 1 L 0.15 M NaCl solution at 4°C to dialyze overnight in the dark.
9. Use 1 L of 0.01 M PBS/ 0.01% sodium azide solution to dialyze at room temperature and avoid light for 4 hours, and then dialyze again overnight at 4°C and avoid light.
10. Filter the antibody solution with a 0.22 μm syringe filter.
11. Dilute the labeled antibody solution with 0.01 M PBS/0.01% sodium azide integer times, and measure the UV-visible absorbance at 280nm (protein) and 552nm (Cy).
12. The product is freeze-dried into powder or stored in 0.01 M PBS/ 0.01% sodium azide solution at -20°C and protected from light.
F/P calculation:
The molar absorption coefficient of Cy3 at 552nm is 150000 M-1cm-1; the molar absorption coefficient of this protein at 280nm is 170 000 M-1cm-1; the molar absorption coefficients of different proteins are different; the absorption of Cy3 dye itself at 280 nm is 8% at 552nm. Calculate the F/P value according to the following formula.
[Cy3] = A552/150000
[antibody] = {A280- (0.08×A552)}/170000
F/P final= [Cy3]/[antibody]= {1.13×A552}/ {A280- (0.08×A552)}
| 储存温度 | 避光,-20°C储存,干燥 |
|---|---|
| 品牌 | Jinpan |
| 产品名称 | Super Fluor 647 SE |
|---|---|
| 英文名称 | Super Fluor 647 SE |
| 运输条件 | 超低温冰袋运输 |
Super Fluor 647 活化酯的性质参数
活化酯分子量:1300
最大吸收波长(nm):647
摩尔吸光系数(M-1cm-1):239000
最大发射波长(nm):665
A280nm /Amax:3%
Super Fluor 标记抗体的参考方案
1) 蛋白溶解：用0.1 M NaHCO3溶解至终浓度为2mg/mL
将1ml 去离子水加入84mgNaHCO3瓶中，配成1M 的碳酸氢钠溶液。振荡或吹打至完全溶解。该溶液pH 约8.3，2～6℃保存可用2 星期；若抗体是溶液，将抗体稀释至2mg/mL，再加入1/10 体积的上述碳酸氢钠溶液；若抗体是冻干粉，将1M 碳酸氢钠溶液用10 倍去离子水稀释成0.1M 后，用其将抗体溶成2mg/mL的溶液；
2）染料溶解：活化Super Flour SE染料固体粉末从冰箱放入干燥器中慢慢升至常温后，用无水DMSO将染料配置浓度为1 mg/mL。
3）标记：在1mL蛋白溶液中缓慢加入适量染料溶液，（蛋白:染料摩尔比=1:20～50；质量比可在以下范围内优化10～200ug Super Flour SE 每毫克IgG抗体）。同时在暗处常温缓慢搅拌1小时。也可以直接将蛋白溶液直接加入染料的固体粉末的试剂瓶中，同时在暗处常温缓慢搅拌1小时。
4）纯化：用透析法或者葡聚糖Sephadex纯化产品后，产品冷冻干燥成粉末或在0.01 M PBS/2mM叠氮化钠溶液中，-20℃避光储存。
5）贮存：标记好的蛋白贮存于2～6℃，避光。若纯化的蛋白偶联物浓度小于1mg/mL，加入BSA 或其他稳定蛋白1～10mg/ml。在2mM 叠氮钠存在的情况下，2～6 度可保存几个月。保存更长时间，则需分装后-20℃保存。避免反复冻融，避光保存。
计算标记比例F/P值
对于IgG 等大多数抗体来说，摩尔吸光系数为203000，F/P值在4～9 之间是最合适。
Super Flour 647标记蛋白F/P值计算：
1）用PBS 精确倍数稀释定量的少许纯化过的偶联蛋白，在1cm比色皿中测定280nm和650nm的光吸收；
2）计算样品中的蛋白浓度：蛋白摩尔浓度=(A280-(A650×0.03))×稀释倍数/203000
3）计算标记比例：每摩尔蛋白结合的染料摩尔数=A650×稀释倍数/(239000×蛋白摩尔浓度)
活性染料稳定性与储存
请注意：未开封的粉末在避光干燥-20℃可存放6月。其水溶液现配现用不能储存。任何溶解后的染料最好立即使用；无水的DMSO溶液-20°C保存最多2个星期。
Properties of Super Fluor 647 activated ester
Activated ester molecular weight: 1300
Maximum absorption wavelength (nm): 647
Molar absorption coefficient (M-1cm-1): 239000
Maximum emission wavelength (nm): 665
A280nm /Amax: 3%
Reference scheme for Super Fluor labeled antibodies
1) Protein dissolution: Dissolve with 0.1 M NaHCO3 to a final concentration of 2mg/mL
Add 1ml of deionized water to an 84mg NaHCO3 bottle to make a 1M sodium bicarbonate solution. Shake or pipette until completely dissolved. The pH of the solution is about 8.3, and it can be stored at 2-6°C for 2 weeks; if the antibody is a solution, dilute the antibody to 2mg/mL, then add 1/10 volume of the above sodium bicarbonate solution; if the antibody is a lyophilized powder, add 1M After the sodium bicarbonate solution is diluted with 10 times deionized water to 0.1M, use it to dissolve the antibody into a 2mg/mL solution;
2) Dye dissolution: After the activated Super Flour SE dye solid powder is slowly warmed to room temperature in a desiccator from the refrigerator, the dye preparation concentration is 1 mg/mL with anhydrous DMSO.
3) Labeling: Slowly add an appropriate amount of dye solution to 1 mL of protein solution, (protein: dye molar ratio = 1:20-50; the mass ratio can be optimized within the following range of 10-200ug Super Flour SE per mg of IgG antibody). At the same time, slowly stir for 1 hour at room temperature in the dark. It is also possible to directly add the protein solution directly to the reagent bottle of the solid powder of the dye, and at the same time, slowly stir for 1 hour at room temperature in the dark.
4) Purification: After purifying the product by dialysis or Sephadex, the product is freeze-dried into powder or stored in 0.01 M PBS/2mM sodium azide solution at -20°C and protected from light.
5) Storage: Store the labeled protein at 2～6℃, away from light. If the concentration of the purified protein conjugate is less than 1 mg/mL, add BSA or other stable proteins at 1-10 mg/ml. In the presence of 2mM sodium azide, it can be stored for several months at 2-6 degrees. For longer storage, you need to store at -20°C after packaging. Avoid repeated freezing and thawing, and store in the dark.
Calculate the mark ratio F/P value
For most antibodies such as IgG, the molar absorption coefficient is 203000, and the F/P value between 4-9 is the most appropriate.
Super Flour 647 labeled protein F/P value calculation:
1) Dilute quantitatively a few purified coupled proteins with PBS with precise multiples, and measure the light absorption at 280nm and 650nm in a 1cm cuvette;
2) Calculate the protein concentration in the sample: protein molar concentration=(A280-(A650×0.03))×dilution multiple/203000
3) Calculate the labeling ratio: moles of dye bound per mole of protein = A650 × dilution factor / (239000 × protein molar concentration)
Reactive dye stability and storage
Please note: the unopened powder can be stored for 6 months in the dark and dry at -20℃. The aqueous solution can not be stored when used. Any dissolved dye is best used immediately; anhydrous DMSO solution is stored at -20°C for up to 2 weeks.
| 储存温度 | 避光,-20°C储存,干燥 |
|---|---|
| 品牌 | Jinpan |
| 产品名称 | Super Fluor 750 SE |
|---|---|
| 英文名称 | Super Fluor 750 SE |
| 运输条件 | 超低温冰袋运输 |
Super Fluor 750 活化酯的性质参数
活化酯分子量:1300
最大吸收波长(nm):750
摩尔吸光系数(M-1cm-1):240000
最大发射波长(nm):775
A280nm /Amax:4%
Super Fluor 标记抗体的参考方案
1) 蛋白溶解：用0.1 M NaHCO3溶解至终浓度为2mg/mL
将1ml 去离子水加入84mgNaHCO3瓶中，配成1M 的碳酸氢钠溶液。振荡或吹打至完全溶解。该溶液pH 约8.3，2～6℃保存可用2 星期；若抗体是溶液，将抗体稀释至2mg/mL，再加入1/10 体积的上述碳酸氢钠溶液；若抗体是冻干粉，将1M 碳酸氢钠溶液用10 倍去离子水稀释成0.1M 后，用其将抗体溶成2mg/mL的溶液。
2）染料溶解：活化Super Flour SE染料固体粉末从冰箱放入干燥器中慢慢升至常温后，用无水DMSO将染料配置浓度为1 mg/mL。
3）标记：在1mL蛋白溶液中缓慢加入适量染料溶液，（蛋白:染料摩尔比=1:20～50；质量比可在以下范围内优化10～200ug Super Flour SE 每毫克IgG抗体）。同时在暗处常温缓慢搅拌1小时。也可以直接将蛋白溶液直接加入染料的固体粉末的试剂瓶中，同时在暗处常温缓慢搅拌1小时。
4）纯化：用透析法或者葡聚糖Sephadex纯化产品后，产品冷冻干燥成粉末或在0.01 M PBS/2mM叠氮化钠溶液中，-20℃避光储存。
5）贮存：标记好的蛋白贮存于2～6℃，避光。若纯化的蛋白偶联物浓度小于1mg/mL，加入BSA 或其他稳定蛋白1～10mg/ml。在2mM 叠氮钠存在的情况下，2～6 度可保存几个月。保存更长时间，则需分装后-20℃保存。避免反复冻融，避光保存。
计算标记比例F/P值
对于IgG 等大多数抗体来说，摩尔吸光系数为203000，F/P值在4～9 之间是最合适。
Super Flour 750标记蛋白F/P值计算：
1）用PBS 精确倍数稀释定量的少许纯化过的偶联蛋白，在1cm比色皿中测定280nm和750nm的光吸收
2）计算样品中的蛋白浓度：蛋白摩尔浓度=(A280-(A750×0.04))×稀释倍数/203000
3）计算标记比例：每摩尔蛋白结合的染料摩尔数=A750×稀释倍数/(240000×蛋白摩尔浓度)
活性染料稳定性与储存
请注意：未开封的粉末在避光干燥-20℃可存放6个月。其水溶液现配现用不能储存。任何溶解后的染料最好立即使用；无水的DMSO溶液-20°C保存最多2个星期。
Properties of Super Fluor 750 activated ester
Activated ester molecular weight: 1300
Maximum absorption wavelength (nm): 750
Molar absorption coefficient (M-1cm-1): 240000
Maximum emission wavelength (nm): 775
A280nm /Amax: 4%
Reference scheme for Super Fluor labeled antibodies
1) Protein dissolution: Dissolve with 0.1 M NaHCO3 to a final concentration of 2mg/mL
Add 1ml of deionized water to an 84mg NaHCO3 bottle to make a 1M sodium bicarbonate solution. Shake or pipette until completely dissolved. The pH of the solution is about 8.3, and it can be stored at 2-6°C for 2 weeks; if the antibody is a solution, dilute the antibody to 2mg/mL, then add 1/10 volume of the above sodium bicarbonate solution; if the antibody is a lyophilized powder, add 1M After the sodium bicarbonate solution is diluted with 10 times deionized water to 0.1M, use it to dissolve the antibody into a 2mg/mL solution.
2) Dye dissolution: After the activated Super Flour SE dye solid powder is slowly warmed to room temperature in a desiccator from the refrigerator, the dye preparation concentration is 1 mg/mL with anhydrous DMSO.
3) Labeling: Slowly add an appropriate amount of dye solution to 1 mL of protein solution, (protein: dye molar ratio = 1:20-50; the mass ratio can be optimized within the following range of 10-200ug Super Flour SE per mg of IgG antibody). At the same time, slowly stir for 1 hour at room temperature in the dark. It is also possible to directly add the protein solution directly to the reagent bottle of the solid powder of the dye, and at the same time, slowly stir for 1 hour at room temperature in the dark.
4) Purification: After purifying the product by dialysis or Sephadex, the product is freeze-dried into powder or stored in 0.01 M PBS/2mM sodium azide solution at -20°C and protected from light.
5) Storage: Store the labeled protein at 2～6℃, away from light. If the concentration of the purified protein conjugate is less than 1 mg/mL, add BSA or other stable proteins at 1-10 mg/ml. In the presence of 2mM sodium azide, it can be stored for several months at 2-6 degrees. For longer storage, you need to store at -20°C after packaging. Avoid repeated freezing and thawing, and store in the dark.
Calculate the mark ratio F/P value
For most antibodies such as IgG, the molar absorption coefficient is 203000, and the F/P value between 4-9 is the most appropriate.
Super Flour 750 labeled protein F/P value calculation:
1) Dilute quantitatively a few purified coupled proteins with PBS with precise multiples, and measure the light absorption at 280nm and 750nm in a 1cm cuvette
2) Calculate the protein concentration in the sample: protein molar concentration=(A280-(A750×0.04))×dilution multiple/203000
3) Calculate the labeling ratio: moles of dye bound per mole of protein = A750 × dilution factor/(240000 × protein molar concentration)
Reactive dye stability and storage
Please note: the unopened powder can be stored for 6 months in the dark and dry at -20℃. The aqueous solution can not be stored when used. Any dissolved dye is best used immediately; anhydrous DMSO solution is stored at -20°C for up to 2 weeks.
| 储存温度 | 避光,-20°C储存,干燥 |
|---|---|
| 品牌 | Jinpan |
| 产品名称 | Cy5 SE |
|---|---|
| 英文名称 | Cy5 SE |
| 运输条件 | 超低温冰袋运输 |
产品简介
菁染料是性能优良的荧光标记染料，摩尔吸光系数在荧光染料中是最高的，其琥珀酰亚胺酯是最常用的脂肪氨基标记试剂，广泛用于蛋白、抗体、核酸及其他生物分子的标记和检测。通过改变次甲基链的长度, 可改变其荧光发射波长，每增加一个双键，按照Huoffman规则正好红移约100nm。
菁染料Cy3和Cy5已成为基因芯片的首选荧光标记物;另外,Cy5, Cy5.5和Cy7的吸收在近红外区背景非常低，是荧光强度最高、最稳定的长波长染料。特别适合于活体小动物体内成像代替放射性元素。
产品参数
Ex(nm) 646 Em(nm) 664 Solvent: Water or DMSO
菁染料琥珀酰亚胺酯（CyDye NHS）的标记
菁染料和生物分子的比例F/P=4～12之间荧光强度最高，F/P值过高荧光探针会自我淬灭并影响生物分子的生物活性，标记生物分子最好是用单琥珀酰亚胺酯，但是用双修饰的CyDye NHS并没有发现交联。CyDye NHS标记抗体在pH (8.5～9.4)时10分钟F/P可达5～6，而在pH 7.0几乎不反应。我们用不同比例的Cy3标记anti-glutathione-S-transferase (GST)多克隆抗体发现用1:1, 5:1, 10:1 和20:1标记时得到的F/P值分别是0.28:1, 1.16:1, 2.3:1 和4.6:1。
操作步骤
水溶性Cy5 NHS标记(D-ser2)-leucineenkephalin
1. Cy5 NHS 1.0 mg 溶解于400 μL DMSO后，加入到1mL 的玻璃瓶中盛有(D-ser2)-leucine-enkephalin acetate (YSGFLT, 0.75 mg) 的400 μL DMSO溶液。（Dye 和peptide 投料比是1:1）
2. 然后加入15 μL三乙胺，常温避光搅拌反应混合物过夜。
3. 用HPLC纯化蛋白，使用蛋白C18柱子(25 cm×10 mm)，每次上样注入2×400 μL，30分钟梯度洗脱从0.1% TFA水溶液到MeCN∶H2O（0.1% TFA）=70∶30，流速4mL /min。（对不同的蛋白选择不同的合适HPLC梯度流动相）
4. 收集适当的色带峰，标记多肽的保留时间比未标记的多肽长。
5. 产品冷冻干燥成粉末或在水溶液中，-20℃避光储存；必要时可用质谱表征。
6. CyDye标记的蛋白稳定性取决于蛋白本身。例如标记的IgG在4 ℃可避光保存2月；更长期的保存需加入等体积的甘油-20 ℃避光保存。
F/P计算
Cy5 在650 nm 的摩尔吸光系数为250000 M-1cm-1，所用蛋白在280 nm处的摩尔吸光系数为170000 M-1cm-1；Cy5在280nm处的吸收是650nm处的5%。按以下公式计算F/P值。
[Cy5 dye] = (A650)/250000
[peptide] =[A280-(0.05×A650)]/170 000
F/P final= [dye]/[peptide]= {0.68×A650}/ {A280- (0.05×A650)}
Product Introduction
Cyanine dye is a fluorescent labeling dye with excellent performance. Its molar absorption coefficient is the highest among fluorescent dyes. Its succinimidyl ester is the most commonly used aliphatic amino group labeling reagent and is widely used for labeling proteins, antibodies, nucleic acids and other biological molecules. And detection. By changing the length of the methine chain, the fluorescence emission wavelength can be changed. For each double bond added, the red shift is exactly 100 nm according to the Huoffman rule.
The cyanine dyes Cy3 and Cy5 have become the first choice fluorescent markers for gene chips; in addition, the absorption of Cy5, Cy5.5 and Cy7 is very low in the near-infrared region, and they are the long-wavelength dyes with the highest fluorescence intensity and the most stable. It is especially suitable for in vivo imaging of small living animals instead of radioactive elements.
Product parameter
Ex(nm) 646 Em(nm) 664 Solvent: Water or DMSO
Labeling of cyanine dye succinimide ester (CyDye NHS)
The ratio of cyanine dye and biomolecule is between F/P=4～12. The fluorescence intensity is the highest. If the F/P value is too high, the fluorescent probe will self-quench and affect the biological activity of biomolecules. It is best to use single amber to label biomolecules. Imide ester, but CyDye NHS modified with double did not find cross-linking. CyDye NHS-labeled antibody can reach 5-6 F/P in 10 minutes at pH (8.5-9.4), but hardly reacts at pH 7.0. We used different ratios of Cy3 to label anti-glutathione-S-transferase (GST) polyclonal antibodies and found that the F/P values obtained when labeled with 1:1, 5:1, 10:1 and 20:1 were 0.28:1 , 1.16:1, 2.3:1 and 4.6:1.
Steps
Water-soluble Cy5 NHS labeling (D-ser2)-leucine enkephalin
1. After dissolving 1.0 mg of Cy5 NHS in 400 μL DMSO, add it to a 1 mL glass bottle containing 400 μL DMSO solution of (D-ser2)-leucine-enkephalin acetate (YSGFLT, 0.75 mg). (The feed ratio of Dye and peptide is 1:1)
2. Then add 15 μL of triethylamine, and stir the reaction mixture overnight at room temperature in the dark.
3. Purify the protein by HPLC, use a protein C18 column (25 cm×10 mm), inject 2×400 μL each time, 30 minutes gradient elution from 0.1% TFA aqueous solution to MeCN: H2O (0.1% TFA) = 70 ：30, flow rate 4mL/min. (Choose different suitable HPLC gradient mobile phases for different proteins)
4. Collect the appropriate color band peaks. The retention time of the labeled peptide is longer than that of the unlabeled peptide.
5. The product is freeze-dried into powder or stored in an aqueous solution at -20°C and protected from light; if necessary, it can be characterized by mass spectrometry.
6. The stability of CyDye-labeled protein depends on the protein itself. For example, labeled IgG can be stored in the dark at 4°C for 2 months; for longer-term storage, an equal volume of glycerol should be added to -20°C in dark storage.
F/P calculation
The molar absorption coefficient of Cy5 at 650 nm is 250 000 M-1cm-1, and the molar absorption coefficient of the protein used at 280 nm is 170 000 M-1cm-1; the absorption of Cy5 at 280 nm is 5% of that at 650 nm. Calculate the F/P value according to the following formula.
[Cy5 dye] = (A650)/250 000
[Peptide] =[A280-(0.05×A650)]/170 000
F/P final= [dye]/[peptide]= {0.68×A650}/ {A280- (0.05×A650)}
| 储存温度 | 避光,-20°C储存,干燥 |
|---|---|
| 品牌 | Jinpan |
| 产品名称 | 6-TET, SE |
|---|---|
| 英文名称 | 6-TET, SE |
| 别名 | 6-Carboxy-2',4,7',7-tetrachlorofluorescein, succinimidyl ester |
| 英文别名 | 6-Carboxy-2′,4,7′,7-tetrachlorofluorescein, succinimidyl ester |
| 运输条件 | 超低温冰袋运输 |
Features and Biological Applications
6-TET, SE is a popular amino-reactive fluorescent probe that is widely used in nucleic acid sequencing and related research.
Ex(nm)：521，Em(nm)：536
References
1. Browne KA. (2005) Sequence-specific, self-reporting hairpin inversion probes. J Am Chem Soc, 127, 1989.
2.Dobson N, McDowell DG, French DJ, Brown LJ, Mellor JM, Brown T. (2003) Synthesis of HyBeacons and dual-labelled probes containing 2′-fluorescent groups for use in genetic analysis. Chem Commun (Camb), 1234.
Features and Biological Applications
6-TET, SE is a popular amino-reactive fluorescent probe that is widely used in nucleic acid sequencing and related research.
Ex(nm)：521，Em(nm)：536
References
1. Browne KA. (2005) Sequence-specific, self-reporting hairpin inversion probes. J Am Chem Soc, 127, 1989.
2.Dobson N, McDowell DG, French DJ, Brown LJ, Mellor JM, Brown T. (2003) Synthesis of HyBeacons and dual-labelled probes containing 2′-fluorescent groups for use in genetic analysis. Chem Commun (Camb), 1234.
| 敏感性 | 对光和湿度敏感 |
|---|---|
| 储存温度 | 避光,-20°C储存,干燥 |
| 品牌 | Jinpan |
| 产品名称 | Super Fluor 555 SE |
|---|---|
| 英文名称 | Super Fluor 555 SE |
| 运输条件 | 超低温冰袋运输 |
Super Fluor 555 活化酯的性质参数
活化酯分子量:1250
最大吸收波长(nm):555
摩尔吸光系数(M-1cm-1):150000
最大发射波长(nm):565
A280nm /Amax:8%
Super Fluor 标记抗体的参考方案
1) 蛋白溶解：用0.1 M NaHCO3溶解至终浓度为2mg/mL
将1ml 去离子水加入84mgNaHCO3瓶中，配成1M 的碳酸氢钠溶液。振荡或吹打至完全溶解。该溶液pH 约8.3，2～6℃保存可用2 星期；若抗体是溶液，将抗体稀释至2mg/mL，再加入1/10 体积的上述碳酸氢钠溶液；若抗体是冻干粉，将1M 碳酸氢钠溶液用10 倍去离子水稀释成0.1M 后，用其将抗体溶成2mg/mL的溶液；
2）染料溶解：活化Super Flour SE染料固体粉末从冰箱放入干燥器中慢慢升至常温后，用无水DMSO将染料配置浓度为1 mg/mL。
3）标记：在1mL蛋白溶液中缓慢加入适量染料溶液，（蛋白:染料摩尔比=1:20～50；质量比可在以下范围内优化10～200ug Super Flour SE 每毫克IgG抗体）。同时在暗处常温缓慢搅拌1小时。也可以直接将蛋白溶液直接加入染料的固体粉末的试剂瓶中，同时在暗处常温缓慢搅拌1小时。
4）纯化：用透析法或者葡聚糖Sephadex纯化产品后，产品冷冻干燥成粉末或在0.01 M PBS/2mM叠氮化钠溶液中，-20℃避光储存。
5）贮存：标记好的蛋白贮存于2～6℃，避光。若纯化的蛋白偶联物浓度小于1mg/mL，加入BSA 或其他稳定蛋白1～10mg/ml。在2mM 叠氮钠存在的情况下，2～6 度可保存几个月。保存更长时间，则需分装后-20℃保存。避免反复冻融，避光保存。
计算标记比例F/P值
对于IgG 等大多数抗体来说，摩尔吸光系数为203000，F/P值在4～9 之间是最合适。
Super Flour 555标记蛋白F/P值计算：
1）用PBS 精确倍数稀释定量的少许纯化过的偶联蛋白，在1cm比色皿中测定280nm和555nm的光吸收；
2）计算样品中的蛋白浓度：蛋白摩尔浓度=(A280-(A555×0.08))×稀释倍数/203000
3）计算标记比例：每摩尔蛋白结合的染料摩尔数=A555×稀释倍数/(150000×蛋白摩尔浓度)
活性染料稳定性与储存
请注意：未开封的粉末在避光干燥-20℃可存放6月。其水溶液现配现用不能储存。任何溶解后的染料最好立即使用；无水的DMSO溶液-20°C保存最多2个星期。
Properties of Super Fluor 555 activated ester
Activated ester molecular weight: 1250
Maximum absorption wavelength (nm): 555
Molar absorption coefficient (M-1cm-1): 150000
Maximum emission wavelength (nm): 565
A280nm / Amax: 8%
Reference scheme for Super Fluor labeled antibodies
1) Protein dissolution: Dissolve with 0.1 M NaHCO3 to a final concentration of 2mg/mL
Add 1ml of deionized water to an 84mg NaHCO3 bottle to make a 1M sodium bicarbonate solution. Shake or pipette until completely dissolved. The pH of the solution is about 8.3, and it can be stored at 2-6°C for 2 weeks; if the antibody is a solution, dilute the antibody to 2mg/mL, then add 1/10 volume of the above sodium bicarbonate solution; if the antibody is a lyophilized powder, add 1M After the sodium bicarbonate solution is diluted with 10 times deionized water to 0.1M, use it to dissolve the antibody into a 2mg/mL solution;
2) Dye dissolution: After the activated Super Flour SE dye solid powder is slowly warmed to room temperature in a desiccator from the refrigerator, the dye preparation concentration is 1 mg/mL with anhydrous DMSO.
3) Labeling: Slowly add an appropriate amount of dye solution to 1 mL of protein solution, (protein: dye molar ratio = 1:20-50; the mass ratio can be optimized within the following range of 10-200ug Super Flour SE per mg of IgG antibody). At the same time, slowly stir for 1 hour at room temperature in the dark. It is also possible to directly add the protein solution directly to the reagent bottle of the solid powder of the dye, and at the same time, slowly stir for 1 hour at room temperature in the dark.
4) Purification: After purifying the product by dialysis or Sephadex, the product is freeze-dried into powder or stored in 0.01 M PBS/2mM sodium azide solution at -20°C and protected from light.
5) Storage: Store the labeled protein at 2～6℃, away from light. If the concentration of the purified protein conjugate is less than 1 mg/mL, add BSA or other stable proteins at 1-10 mg/ml. In the presence of 2mM sodium azide, it can be stored for several months at 2-6 degrees. For longer storage, you need to store at -20°C after packaging. Avoid repeated freezing and thawing, and store in the dark.
Calculate the mark ratio F/P value
For most antibodies such as IgG, the molar absorption coefficient is 203000, and the F/P value between 4-9 is the most appropriate.
Super Flour 555 labeled protein F/P value calculation
1) Dilute quantitatively a few purified coupled proteins with PBS with precise multiples, and measure the light absorption at 280nm and 555nm in a 1cm cuvette;
2) Calculate the protein concentration in the sample: protein molar concentration=(A280-(A555×0.08))×dilution multiple/203000
3) Calculate the labeling ratio: moles of dye bound per mole of protein = A555 × dilution factor / (150000 × protein molar concentration)
Reactive dye stability and storage
Please note: the unopened powder can be stored for 6 months in the dark and dry at -20℃. The aqueous solution can not be stored when used. Any dissolved dye is best used immediately; anhydrous DMSO solution is stored at -20°C for up to 2 weeks.
| 储存温度 | 避光,-20°C储存,干燥 |
|---|---|
| 品牌 | Jinpan |
| 产品名称 | Cy7 SE |
|---|---|
| 英文名称 | Cy7 SE |
| 别名 | Cy7 NHS |
| 英文别名 | Cy7 NHS |
| 运输条件 | 超低温冰袋运输 |
产品简介
水溶性菁染料是性能优良的荧光标记染料, 具有很好的水溶性和很高的摩尔吸光系数，其琥珀酰亚胺酯是最常用的脂肪氨基标记试剂，广泛用于蛋白、抗体、核酸及其他生物分子的标记和检测。特别是Cy5.5和Cy7的吸收在近红外区，已广泛用于活体成像领域来代替放射性元素。在哺乳动物体内血红蛋白是吸收可见光的主要成分, 能吸收中蓝绿光波段的大部分可见光; 水和脂质主要吸收红外线, 因此选择激发和发射光谱位于650nm-900nm(近红外窗口)的近红外荧光标记的红光虽然有部分散射消耗但大部分可以穿透哺乳动物组织被敏感的CCD检测到，更有利于活体光学成像，特别是深层组织的荧光成像。
产品参数
Ex(nm)：747
Em(nm)：774
Solvent：Water or DMSO
裸鼠的活体成像实例
SPF级BALB/C裸鼠，6-8 周龄, 18-20克, 实验前24 h 自由进食、饮水。
实验裸鼠麻醉后，将裸鼠俯卧位平放于小动物多光谱活体成像系统的记录暗箱中。实验时将Cy7或Cy7标记的生物分子或药物DMSO稀释后,于裸鼠尾静脉注射200μL( 0.5 mg/g) [最佳用量和时间需要客户根据自己的仪器和药物试剂等条件优化],每5min 记录1 张动物在体内发射荧光的成像图片,分析荧光药物的分布情况。对照鼠不注射药物，进行同时记录。记录结束后迅速解剖裸鼠的心、肝、脾、肺、肾等脏器，进行成像。
Cy7 检测时激发波长700～770 nm带通,发射波长790 nm长通。液晶可调谐滤光片扫描范围780～950 nm ,扫描步进10 nm。曝光时间为500 ms。
不同的药物代谢时间不一样，注射入裸鼠体内，荧光立即分布全身，然后逐步向膀胱聚集，呈现显著的肾排泄的特点一般4~6小时，快得只有30分钟；如果是骨骼等部位靶点的Cy7标记药物，有客户反映一周后活体成像系统仍能检测到荧光成像。
器官切片观察：将解剖的器官迅速放置于4%多聚甲醛固定4小时以上，0%，20%，30%PBS蔗糖依次沉底，20μm切片，多聚赖氨酸洗过的载玻片贴片，晾干，DAPI染色。共聚焦显微镜观察，激光器为氦氖633 nm。
稳定性与储存
未开封的粉末在避光干燥-20℃存放12月。CyDye NHS水溶液现配现用不能储存。任何溶解后的CyDye NHS粉末最好立即使用；无水的DMSO溶液-20°C保存最多2个星期。
Product introduction
Water soluble cyanine dye is a kind of fluorescent labeling dye with excellent performance. It has good water solubility and high molar absorptivity. Its succinimide ester is the most commonly used fatty amino labeling reagent, which is widely used for the labeling and detection of proteins, antibodies, nucleic acids and other biological molecules. In particular, the absorption of cy5.5 and cy7 in the near infrared region has been widely used in the field of in vivo imaging to replace radioactive elements. In mammals, hemoglobin is the main component of absorption of visible light, which can absorb most visible light in the blue-green band; water and lipid mainly absorb infrared light, Therefore, although the red light with excitation and emission spectrum located at 650 nm-900 nm (near-infrared window) has some scattering consumption, most of the red light can penetrate mammalian tissues and be detected by sensitive CCD, which is more conducive to in vivo optical imaging, especially fluorescence imaging of deep tissue.
Product parameters
Ex(nm)：747
Em(nm)：774
Solvent：Water or DMSO
Living imaging of nude mice
SPF BALB / C nude mice, 6-8 weeks old, 18-20g, were free to eat and drink 24 hours before the experiment.
After anesthetized, the nude mice were placed in prone position in the recording box of small animal multispectral imaging system. In the experiment, cy7 or cy7 labeled biomolecule or drug DMSO was diluted, and then 200 μ L (0.5 mg / g) was injected into the tail vein of nude mice [the optimal dosage and time need to be optimized according to the conditions of their own instruments and drug reagents], and 1 imaging picture of the animal emission in vivo was recorded every 5 minutes to analyze the distribution of fluorescent drugs. The control rats were not injected with drugs, and were recorded at the same time. After recording, the heart, liver, spleen, lung, kidney and other organs of nude mice were dissected and imaged.
The excitation wavelength is 700-770 nm and the emission wavelength is 790 nm. The scanning range of liquid crystal tunable filter is 780-950 nm and the scanning step is 10 nm. The exposure time was 500 ms.
Different drug metabolism time is different. When injected into nude mice, fluorescence immediately distributes throughout the body, and then gradually accumulates to the bladder, showing the characteristics of significant renal excretion, generally 4-6 hours, as fast as 30 minutes; if it is cy7 labeled drugs targeted at bone and other parts, some customers report that fluorescence imaging can still be detected in vivo imaging system one week later.
Observation of organ sections: the dissected organs were quickly placed in 4% paraformaldehyde fixed for more than 4 hours, 0%, 20%, 30% PBS sucrose were successively sunk to the bottom, 20 μ m slices were sliced, the slides were pasted with polylysine acid, dried and DAPI staining. The laser is 633 nm He Ne.
Stability and storage
The unsealed powder was dried away from light and stored at – 20 ℃ for 12 months. CyDye NHS aqueous solution can not be stored when it is prepared and used. Any dissolved CyDye NHS powder should be used immediately; the anhydrous DMSO solution should be stored at – 20 ° C for up to 2 weeks.
| 储存温度 | 避光,-20°C储存,干燥 |
|---|---|
| 品牌 | Jinpan |
| 产品名称 | Super Fluor 680 SE |
|---|---|
| 英文名称 | Super Fluor 680 SE |
| 运输条件 | 超低温冰袋运输 |
Super Fluor 680活化酯的性质参数
活化酯分子量:1150
最大吸收波长(nm):680
摩尔吸光系数(M-1cm-1):184000
最大发射波长(nm):702
A280nm /Amax:5%
Super Fluor 标记抗体的参考方案
1) 蛋白溶解：用0.1 M NaHCO3溶解至终浓度为2mg/mL
将1ml 去离子水加入84mgNaHCO3瓶中，配成1M 的碳酸氢钠溶液。振荡或吹打至完全溶解。该溶液pH 约8.3，2～6℃保存可用2 星期；若抗体是溶液，将抗体稀释至2mg/mL，再加入1/10 体积的上述碳酸氢钠溶液；若抗体是冻干粉，将1M 碳酸氢钠溶液用10 倍去离子水稀释成0.1M 后，用其将抗体溶成2mg/mL的溶液；
2）染料溶解：活化Super Flour SE染料固体粉末从冰箱放入干燥器中慢慢升至常温后，用无水DMSO将染料配置浓度为1 mg/mL。
3）标记：在1mL蛋白溶液中缓慢加入适量染料溶液，（蛋白:染料摩尔比=1:20～50；质量比可在以下范围内优化10～200ug Super Flour SE 每毫克IgG抗体）。同时在暗处常温缓慢搅拌1小时。也可以直接将蛋白溶液直接加入染料的固体粉末的试剂瓶中，同时在暗处常温缓慢搅拌1小时。
4）纯化：用透析法或者葡聚糖Sephadex纯化产品后，产品冷冻干燥成粉末或在0.01 M PBS/2mM叠氮化钠溶液中，-20℃避光储存。
5）贮存：标记好的蛋白贮存于2～6℃，避光。若纯化的蛋白偶联物浓度小于1mg/mL，加入BSA 或其他稳定蛋白1～10mg/ml。在2mM 叠氮钠存在的情况下，2～6 度可保存几个月。保存更长时间，则需分装后-20℃保存。避免反复冻融，避光保存。
计算标记比例F/P值
对于IgG 等大多数抗体来说，摩尔吸光系数为203000，F/P值在4～9 之间是最合适。
Super Flour 680标记蛋白F/P值计算：
1）用PBS 精确倍数稀释定量的少许纯化过的偶联蛋白，在1cm比色皿中测定280nm和680nm的光吸收
2）计算样品中的蛋白浓度：蛋白摩尔浓度=(A280-(A650×0.05))×稀释倍数/203000
3）计算标记比例：每摩尔蛋白结合的染料摩尔数=A650×稀释倍数/(184000×蛋白摩尔浓度)
活性染料稳定性与储存
请注意：未开封的粉末在避光干燥-20℃可存放6个月。其水溶液现配现用不能储存。任何溶解后的染料最好立即使用；无水的DMSO溶液-20°C保存最多2个星期。
Properties of Super Fluor 680 activated ester
Activated ester molecular weight: 1150
Maximum absorption wavelength (nm): 680
Molar absorption coefficient (M-1cm-1): 184000
Maximum emission wavelength (nm): 702
A280nm / Amax: 5%
Reference scheme for Super Fluor labeled antibodies
1) Protein dissolution: Dissolve with 0.1 M NaHCO3 to a final concentration of 2mg/mL
Add 1ml of deionized water to an 84mg NaHCO3 bottle to make a 1M sodium bicarbonate solution. Shake or pipette until completely dissolved. The pH of the solution is about 8.3, and it can be stored at 2-6°C for 2 weeks; if the antibody is a solution, dilute the antibody to 2mg/mL, then add 1/10 volume of the above sodium bicarbonate solution; if the antibody is a lyophilized powder, add 1M After the sodium bicarbonate solution is diluted with 10 times deionized water to 0.1M, use it to dissolve the antibody into a 2mg/mL solution;
2) Dye dissolution: After the activated Super Flour SE dye solid powder is slowly warmed to room temperature in a desiccator from the refrigerator, the dye preparation concentration is 1 mg/mL with anhydrous DMSO.
3) Labeling: Slowly add an appropriate amount of dye solution to 1 mL of protein solution, (protein: dye molar ratio = 1:20-50; the mass ratio can be optimized within the following range of 10-200ug Super Flour SE per mg of IgG antibody). At the same time, slowly stir for 1 hour at room temperature in the dark. It is also possible to directly add the protein solution directly to the reagent bottle of the solid powder of the dye, and at the same time, slowly stir for 1 hour at room temperature in the dark.
4) Purification: After purifying the product by dialysis or Sephadex, the product is freeze-dried into powder or stored in 0.01 M PBS/2mM sodium azide solution at -20°C and protected from light.
5) Storage: Store the labeled protein at 2～6℃, away from light. If the concentration of the purified protein conjugate is less than 1 mg/mL, add BSA or other stable proteins at 1-10 mg/ml. In the presence of 2mM sodium azide, it can be stored for several months at 2-6 degrees. For longer storage, you need to store at -20°C after packaging. Avoid repeated freezing and thawing, and store in the dark.
Calculate the mark ratio F/P value
For most antibodies such as IgG, the molar absorption coefficient is 203000, and the F/P value between 4-9 is the most appropriate.
Super Flour 680 labeled protein F/P value calculation:
1) Dilute quantitatively a few purified coupled proteins with PBS with precise multiples, and measure the light absorption at 280nm and 680nm in a 1cm cuvette
2) Calculate the protein concentration in the sample: protein molar concentration=(A280-(A650×0.05))×dilution multiple/203000
3) Calculate the labeling ratio: moles of dye bound per mole of protein = A650 × dilution factor / (184000 × protein molar concentration)
Reactive dye stability and storage
Please note: the unopened powder can be stored for 6 months in the dark and dry at -20℃. The aqueous solution can not be stored when used. Any dissolved dye is best used immediately; anhydrous DMSO solution is stored at -20°C for up to 2 weeks.
| 储存温度 | 避光,-20°C储存,干燥 |
|---|---|
| 品牌 | Jinpan |
| 产品名称 | 铬蓝SE |
|---|---|
| 英文名称 | Eriochrome blue SE |
| 别名 | 铬蓝SE;媒介蓝13;3-[(5-氯-2-羟基苯基)偶氮]-4,5-二羟基-2,7-萘二磺酸二钠盐 |
| 英文别名 | Mordant Blue 13 Plasmocorinth B C.I. 16680 Sunchromine Fast Blue MB |
| 规格或纯度 | 高纯级,Dye content 80% |
溶于水呈品红色，溶于溶纤素，稍溶于乙醇。在浓硫酸中呈溶蓝色，稀释后呈品红色、在浓硝酸中呈棕光红色溶注估烧碱液中呈红光蓝色溶液。其水溶液，加浓盐酸呈品红色、加浓氢氧化钠呈蓝色。染色时遇铜、铁，色光变化大。水中溶解度（80℃）50g/L。
溶于水呈品红色，溶于溶纤素，稍溶于乙醇。在浓硫酸中呈溶蓝色，稀释后呈品红色、在浓硝酸中呈棕光红色溶注估烧碱液中呈红光蓝色溶液。其水溶液，加浓盐酸呈品红色、加浓氢氧化钠呈蓝色。染色时遇铜、铁，色光变化大。水中溶解度（80℃）50g/L。
| 个人防护设备 | Eyeshields,Gloves,type N95 (US),type P1 (EN143) respirator filter |
|---|---|
| WGK | 3 |
| CAS编号 | 1058-92-0 |
|---|---|
| 储存温度 | 密封阴凉干燥保存。 |
| MDL号 | MFCD00003941 |
| 密度 | 1.2870 |
| 分子量 | 518.81 |
| 分子式 | C16H9ClN2Na2O9S2 |
| EC号 | 213-894-4 |
| 品牌 | Jinpan |
| Smiles | [Na+].[Na+].Clc1ccc(O)c(/N=N/c2c(O)c3c(O)cc(cc3cc2S(=O)(=O)[O-])S(=O)(=O)[O-])c1;[Na+].[Na+].Clc1ccc(O)c(N/N=C2/C(=O)c3c(O)cc(cc3C=C2S(=O)(=O)[O-])S(=O)(=O)[O-])c1 |
| PubChem CID | 13991 |
