All lanes : Anti-p38 beta/MAPK11 + p38 alpha/MAPK14 antibody [Y122] (ab32142) at 1/1000 dilution

Lane 1 : Recombinant Human p38 beta/MAPK11 protein (ab117219)
Lane 2 : Recombinant Human p38 gamma/MAPK12 protein (ab117221)
Lane 3 : Recombinant Human p38 delta/MAPK13 protein (ab113869)
Lane 4 : Recombinant human p38 alpha/MAPK14 protein (ab82188)

Predicted band size: 41 kDa

All lanes : Anti-p38 beta/MAPK11 + p38 alpha/MAPK14 antibody [Y122] (ab32142)

Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : Wild-type HEK-293T cell lysate
Lane 4 : MAPK14 knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 41 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab32142 observed at 40 kDa. Red - loading control, ab130007 observed at 125 kDa.  

 ab32142 was shown to react with p38 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255406 (knockout cell lysate ab263787) was used. Wild-type and p38 knockout samples were subjected to SDS-PAGE. ab32142 and Anti-Vinculin antibody [VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and 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 : Anti-p38 beta/MAPK11 + p38 alpha/MAPK14 antibody [Y122] (ab32142)

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : p38 knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 41 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab32142 observed at 40 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab32142 was shown to specifically react with p38 when p38 knockout samples were used. Wild-type and p38 knockout samples were subjected to SDS-PAGE. ab32142 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and 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.

Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) cells labelling p38 (green) with purified ab32142 at 1/250. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).

Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

Ab32142, at a 1/100 dilution, staining p38 in paraffin embedded human skin carcinoma tissue by immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

All lanes : Anti-p38 beta/MAPK11 + p38 alpha/MAPK14 antibody [Y122] (ab32142)

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : p38 knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 41 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab32142 observed at 40 kDa. Red - loading control, ab8245, observed at 37 kDa.

This western blot image is a comparison between ab32142 and a competitor's top cited rabbit polyclonal antibody.

All lanes : Anti-p38 beta/MAPK11 + p38 alpha/MAPK14 antibody [Y122] (ab32142) at 1/50000 dilution

Lane 1 : Hela cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : K562 cell lysate
Lane 4 : MCF7 cell lysate

Predicted band size: 41 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?



10 ug protein per lane