Calreticulin Polyclonal Antibody for Western Blot, IF, ICC, Flow and IP
Calreticulin Polyclonal Antibody for Western Blot, IF, ICC, Flow and IP
Immunofluorescent analysis of Calreticulin (green) U2OS cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS 0.1% triton-X (Product #37525) for 30 minutes at room temperature. Cells were probed with a rabbit polyclonal antibody recognizing Calreticulin (Product # PA3-900) at a dilution of 1:50 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with DyLight 488 goat-anti-rabbit IgG secondary antibody (Product #35552) at a dilution of 1:250 for 30 minutes at room temperature. Actin filaments (red) were stained with DyLight 554-Phalloidin (Product# 21834) at a dilution of 1:300 (i.e. 1unit/mL final concentration) in PBS and incubated for 30 min including. Nuclei (blue) were stained with Hoechst 33342 dye (Product# 62249) at a dilution of 1µg/mL. Images were taken on a Thermo Scientific ArrayScan VTI at 20X magnification.
Immunofluorescent analysis of Beta-Tubulin (red) in HEK293T cells. Cells fixed in 4% formaldehyde were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with a Beta-Tubulin monoclonal antibody (Product # MA5-16308) at a dilution of 1:100 overnight at 4°C in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with a fluorophore-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a Leica DM1000 microscope at 40X magnification. Data courtesy of the Innovators Program.
Immunofluorescent analysis of SOX2 using anti-SOX2 polyclonal antibody (Product# PA1-094) shows specific expression in human embryonal carcinoma NTERA-2 cells (shown in green) but not in negative control HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product #37525) for 15 minutes at room temperature. Cells were probed with a rabbit polyclonal antibody recognizing SOX2 (Product# PA1-094), at a dilution of 1:200 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with DyLight 488 goat-anti-rabbit IgG secondary antibody (Product# 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DY-547 phalloidin, nuclei (blue) were stained with Hoechst 33342 dye (Product# 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
Immunofluorescent analysis of GAPDH (green) in HeLa. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 5% normal goat serum (Product # 31873) for 15 minutes at room temperature. Cells were probed with a GAPDH monoclonal antibody (Product # MA5-15738) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.
Immunofluorescent analysis of Lin28 (red) in H9 embryonic stem cells grown for a few days on Matrigel-coated chamber slides. Cells fixed in 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were probed with a Lin28 polyclonal antibody (Product # PA1-096) at a dilution of 1:200 overnight at 4°C, washed with PBST, and incubated with a fluorescently-conjugated secondary antibody at a dilution of 1:100 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI and cells were analyzed by fluorescence microscopy at 20X magnification.
Immunofluorescent analysis of Sox2 (green) in H9 embryonic stem cells grown for a few days on Matrigel-coated chamber slides. Cells fixed in 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were probed with a Sox2 monoclonal antibody (Product # MA1-014) at a dilution of 1:200 overnight at 4°C, washed with PBST, and incubated with a fluorescein-conjugated secondary antibody at a dilution of 1:100 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI and cells were analyzed by fluorescence microscopy at 20X magnification.
Immunofluorescent analysis of Phalloidin (magenta) and HDAC2 (green) in A431 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA (Product # 37525) in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a HDAC2 polyclonal antibody (Product # PA1-861) at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 633 Phalloidin (Product # 21840) at a dilution of 1:120 (2.5units/ml final concentration) and nuclei (blue) were stained with Hoechst (Product # 62249) at a concentration of 1ug/ml for 30 minutes. Images were taken on a Zeiss Axio Observer Z1 microscope at 20X magnification.
Immunofluorescent analysis of Nestin (green) in human neural stem cells derived from PD-3 iPSCs using Gibco® PSC Neural Induction Medium (Product # A1647801). The cells were fixed and permeabilized using Image-IT® Fixation/Permeabilization kit (Product # R37602), and blocked with the including blocking buffer for one hour at room temperature. Cells were stained with a Nestin monoclonal antibody (Product # MA1-110) at a dilution of 1:100 in blocking buffer for 3 hours at room temperature, and then incubated with a DyLight488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:250 for 1 hour at room temperature. Nucleus DNA (blue) was stained with DAPI (Product # D1306). Images were taken on an EVOS® FLoid® Cell Imaging Station at 10X magnification.
Immunofluorescence analysis of Phospho-P38 pThr180 / pTyr182 Antibody was done on 70% confluent log phase SHSY5Y cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Phospho-P38 pThr180 / pTyr182 Antibody (44684g) at 1µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa flour 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.
Immunofluorescence analysis of iNOS was performed using 70% confluent log phase A549 cells treated with a combination of TNF-α (5 ng/ml), IFNγ (50 U/ml) and IL-1β (50 U/ml) for 18 h. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with iNOS Rabbit Polyclonal Antibody (PA3030A) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e shows the untreated cells with no signal. Panel f represents the no primary control. The images were captured at 60X magnification.