Anti-IKK alpha antibody [Y463] - BSA and Azide free

7 product images

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  Immunoprecipitation - Anti-IKK alpha antibody [Y463] - BSA and Azide free (ab169743)

  Purified Anti-IKK alpha antibody [Y463] ab32041 at 1/60 dilution (2μg) immunoprecipitating IKK alpha in HeLa whole cell lysate.
  Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
  Lane 2 (+): Anti-IKK alpha antibody [Y463] ab32041 + HeLa whole cell lysate.
  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab169743 in HeLa whole cell lysate.
  VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) (1/1000 dilution) was used for Western blotting.
  Blocking Buffer and concentration: 5% NFDM/TBST.
  Diluting buffer and concentration: 5% NFDM/TBST.
  Observed band size: 88 kDa
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-IKK alpha antibody [Y463] ab32041).

  All lanes: Immunoprecipitation - Anti-IKK alpha antibody [Y463] (Anti-IKK alpha antibody [Y463] ab32041)

  Predicted band size: 84 kDa

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  Western blot - Anti-IKK alpha antibody [Y463] - BSA and Azide free (ab169743)

  All lanes: Western blot - Anti-IKK alpha antibody [Y463] (Anti-IKK alpha antibody [Y463] ab32041) at 1/1000 dilution

  Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 15 µg

  Lane 2: Mouse kidney lysate at 15 µg

  Lane 3: C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg

  Lane 4: Rat kidney lysate 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: 84 kDa

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  Western blot - Anti-IKK alpha antibody [Y463] - BSA and Azide free (ab169743)

  All lanes: Western blot - Anti-IKK alpha antibody [Y463] (Anti-IKK alpha antibody [Y463] ab32041) at 1/10000 dilution

  All lanes: Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysate

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

  Predicted band size: 84 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [Y463] - BSA and Azide free (ab169743)

  This data was developed using Anti-IKK alpha antibody [Y463] ab32041, the same antibody clone in a different buffer formulation.
  
  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian cancer tissue sections labeling IKK alpha with purified Anti-IKK alpha antibody [Y463] ab32041 at 1/50 dilution (3.92 μg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

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  Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [Y463] - BSA and Azide free (ab169743)

  Overlay histogram showing HAP1 wildtype (green line) and HAP1-CHUK knockout cells (red line) stained with Anti-IKK alpha antibody [Y463] ab32041. The cells were fixed with 4% formaldehyde (10 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 (Anti-IKK alpha antibody [Y463] ab32041, 1μg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) presorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG isotype control antibody (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-CHUK knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-IKK alpha antibody [Y463] ab32041).

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  Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [Y463] - BSA and Azide free (ab169743)

  This data was developed using Anti-IKK alpha antibody [Y463] ab32041, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of Daudi (Human Burkitt's lymphoma lymphoblast) cells labelling IKK alpha with purified Anti-IKK alpha antibody [Y463] ab32041 at 1/20 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [Y463] - BSA and Azide free (ab169743)

  This data was developed using Anti-IKK alpha antibody [Y463] ab32041, the same antibody clone in a different buffer formulation.
  
  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling IKK alpha with purified Anti-IKK alpha antibody [Y463] ab32041 at 1/50 dilution (3.92 μg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.