Anti-Bax antibody [E63] - BSA and Azide free

9 product images

• 

  Immunoprecipitation - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

  Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, 10μg
  Lane 2 (+): HeLa whole cell lysate
  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Bax antibody [E63] ab32503 in HeLa whole cell lysate
  

  Purified Anti-Bax antibody [E63] ab32503 immunoprecipitating Bax in HeLa lysates. For western blotting, the primary antibody used was purified Anti-Bax antibody [E63] ab32503 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) ab131366 VeriBlot for IP (HRP) was used for detection at 1/1000 dilution. Capture antibody was used at a 1/20 dilution. Blocking and diluting buffer used was 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-Bax antibody [E63] ab32503).

  All lanes: Immunoprecipitation - Anti-Bax antibody [E63] (Anti-Bax antibody [E63] ab32503)

  Predicted band size: 21 kDa

  Observed band size: 21 kDa

  Exposure time: 1s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

  Purified Anti-Bax antibody [E63] ab32503 staining Bax in Human lung carcinoma tissue section by immmunohistochemistry (IHC-P- Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraffin and heat mediated antigen retrieval was performed using EDTA buffer (pH 9.0). Samples were incubated with primary antibody at 1:500 dilution. A goat anti-rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) was used as a secondary antibody at 1:500 dilution. Cytoplasmic staining on human lung carcinoma.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Bax antibody [E63] ab32503).

• 

  Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Bax antibody [E63] ab32503).
  

  Lanes 1- 2: Merged signal (red and green). Green - Anti-Bax antibody [E63] ab32503 observed at 21 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

  Anti-Bax antibody [E63] ab32503 was shown to react with Bax in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human BAX knockout HeLa cell line ab255363 (knockout cell lysate Human BAX knockout HeLa cell lysate ab263841) was used. Wild-type HeLa and BAX knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-Bax antibody [E63] ab32503 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-Bax antibody [E63] (Anti-Bax antibody [E63] ab32503) at 1/1000 dilution

  Lane 1: Wild-type HeLa cell lysate at 20 µg

  Lane 2: BAX knockout HeLa cell lysate at 20 µg

  Lane 2: Western blot - Human BAX knockout HeLa cell line (Human BAX knockout HeLa cell line ab255363)

  Performed under reducing conditions.

  Predicted band size: 21 kDa

  Observed band size: 21 kDa

• 

  Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Bax antibody [E63] ab32503).
  

  Lanes 1 - 2: Merged signal (red and green). Green - Anti-Bax antibody [E63] ab32503 observed at 20 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

  
  Anti-Bax antibody [E63] ab32503 was shown to recognize Bax in wild-type HAP1 cells, along with additional cross-reactive bands. Wild-type and Bax knockout samples were subjected to SDS-PAGE. Anti-Bax antibody [E63] ab32503 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  All lanes: Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985) at 1/1000 dilution

  Lane 1: Wild-type HAP1 cell lysate at 20 µg

  Lane 2: BAX knockout HAP1 cell lysate at 20 µg

  Predicted band size: 21 kDa

• 

  Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

  This data was developed using Anti-Bax antibody [E63] ab32503, the same antibody clone in a different buffer formulation. Different batches of Anti-Bax antibody [E63] ab32503 were tested on HeLa (Human cervix adenocarcinoma epithelial cell) lysate at 0.1 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 18 kDa.

  All lanes: Western blot - Anti-Bax antibody [E63] (Anti-Bax antibody [E63] ab32503)

  Predicted band size: 21 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

  IHC image of Anti-Bax antibody [E63] ab32503 staining Bax in rat kidney formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with Anti-Bax antibody [E63] ab32503, 1:250 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Bax antibody [E63] ab32503).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

  Immunohistochemical analysis of paraffin-embedded human lymph node using anti-Bax Rabbit Monoclonal Antibody (Anti-Bax antibody [E63] ab32503) at 1/250 dilution.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Bax antibody [E63] ab32503).

  Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• 

  Sandwich ELISA - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

  Standard Curve for Bax (Analyte: Recombinant human Bax protein (tagged) ab85157) dilution range 1pg/ml to 1ug/ml using Capture Antibody Mouse monoclonal [2D2] to Bax - BSA and Azide free (ab77566) at 0.2ug/ml and Detector Antibody Rabbit monoclonal [E63] to Bax (Anti-Bax antibody [E63] ab32503) at 0.5ug/ml. Concentration of Anti-Bax antibody [E63] ab32503 may vary from lot to lot; please use this curve as guideline.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Bax antibody [E63] ab32503).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

  This data was produced using the same antibody clone but in a different formulation containing PBS, sodium azide, glycerol and BSA (Anti-Bax antibody [E63] ab32503).

  Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling BAX with Anti-Bax antibody [E63] ab32503 at a concentration of 0.05 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

  Anti-Bax antibody [E63] ab32503 Anti-Bax antibody [E63] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

  Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)