Anti-GAPDH antibody [EPR16891] - BSA and Azide free

11 product images

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  Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  GAPDH was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 at 1/60 dilution. Western blot was performed from the immunoprecipitate using Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract.

  Lane 2: PBS instead of HeLa whole cell extract.

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602).

  All lanes: Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602)

  Predicted band size: 36 kDa

  Observed band size: 36 kDa

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  Western blot - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  Blocking and Diluting buffer and concentration:5% NFDM/TBST
  

  Exposure time:3 seconds

  All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553) at 1/100000 dilution

  All lanes: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 36 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  Immunocytochemistry/immunofluorescence staining of 4% paraformaldehyde fixed; 0.1% triton X 100 permeabilized HeLa (human cervix adenocarcinoma) cells labeling GAPDH with Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 at dilution of 1/500. The secondary antibody used was Alexa Fluor^® 488; goat anti-rabbit IgG (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400. Nucleus was counter-stained with DAPI (blue). Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/500) was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor^®594 goat anti-mouse secondary, 1/500). The negative controls are shown in the bottom middle and right hand panels- for negative control 1 primary antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602; 1/500) and secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120; 1/500) was used. For negative control 2 primary antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291; 1/500) and secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077; 1/400) was used.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling GAPDH with Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocyte of rat spleen is observed. Counter stained with Hematoxylin.
  

  Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602).

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling GAPDH with Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 at 1/180 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602).

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  Western blot - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  This data was developed using Anti-GAPDH antibody [EPR16891] - Loading Control ab181602, the same antibody clone in a different buffer formulation. Different batches of Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 were tested on HeLa (Human cervix adenocarcinoma epithelial cell) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 36 kDa.

  All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602)

  Predicted band size: 36 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  Clone EPR16891 (ab199553) has been successfully conjugated by Abcam. This image was generated using Anti-GAPDH antibody [EPR16891] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-GAPDH antibody [EPR16891] ab201768 for protocol details.

  Alexa Fluor® 488 Anti-GAPDH antibody [EPR16891] ab201768 staining GAPDH in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Alexa Fluor® 488 Anti-GAPDH antibody [EPR16891] ab201768 at 1/200 dilution (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling GAPDH with Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic and nucleus staining on the tumor cells of transitional cell carcinoma of human bladder is observed. Counter stained with Hematoxylin.
  

  Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602).

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  Clone EPR16891 (ab199553) has been successfully conjugated by Abcam. This image was generated using Anti-GAPDH antibody [EPR16891] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-GAPDH antibody [EPR16891] ab201272 for protocol details.

  Alexa Fluor® 647 Anti-GAPDH antibody [EPR16891] ab201272 staining GAPDH in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Alexa Fluor® 647 Anti-GAPDH antibody [EPR16891] ab201272 at 1/100 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 2µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  Clone EPR16891 (ab199553) has been successfully conjugated by Abcam. This image was generated using Anti-GAPDH antibody [EPR16891] (PE). Please refer to PE Anti-GAPDH antibody [EPR16891] ab224004 for protocol details.
  

  Overlay histogram showing HeLa cells stained with PE Anti-GAPDH antibody [EPR16891] ab224004 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (PE Anti-GAPDH antibody [EPR16891] ab224004, 1/1000 dilution) for 30 min at 22°C.

  Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

  Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

  This antibody gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)

  Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling GAPDH with Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.
  

  Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602).

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.