Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138]

9 product images

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  Western blot - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

  Lanes 1 - 4: Green - ab31828 observed at 43 kDa.
  

  ab31828 was shown to react with Anti-p38 antibody [M138] in Western blot. Membranes were blocked in 100% Licor before incubation with ab31828 and overnight at 4 °C at a 1 in 1000 dilution. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) secondary antibody at 1 in 20000 dilution for 1 h at room temperature before imaging.

  All lanes: Western blot - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828) at 1/1000 dilution

  Lane 1: MAPK11 recombinant (Recombinant Human p38 beta/MAPK11 protein ab117219) at 0.5 µg

  Lane 2: MAPK12 recombinant(ab 117221) at 0.5 µg

  Lane 3: MAPK13 recombinant (Recombinant Human p38 delta/MAPK13 protein ab113869) at 0.5 µg

  Lane 4: MAPK14 (p38) recombinant (Recombinant Human p38 alpha/MAPK14 protein ab82188) at 0.5 µg

  Performed under reducing conditions.

  Observed band size: 43 kDa

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  Western blot - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

  All lanes: Western blot - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828) at 1/1000 dilution

  Lane 1: Western blot - Recombinant Human p38 beta/MAPK11 protein (Recombinant Human p38 beta/MAPK11 protein ab117219)

  Lane 2: Western blot - Recombinant Human p38 gamma/MAPK12 protein (Recombinant Human p38 gamma/MAPK12 protein ab117221)

  Lane 3: Western blot - Recombinant Human p38 delta/MAPK13 protein (Recombinant Human p38 delta/MAPK13 protein ab113869)

  Lane 4: Western blot - Recombinant Human p38 alpha/MAPK14 protein (Recombinant Human p38 alpha/MAPK14 protein ab82188)

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

  IHC image of ab31828 staining p38 in normal human esophagus 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 ab31828, 1/50 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 negative 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.

  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

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  Western blot - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

  Lane 1: Wild-type HAP1 cell lysate (20 μg)
  Lane 2: p38 knockout HAP1 cell lysate (20 μg)
  Lane 3: HeLa cell lysate (20 μg)
  Lane 4: Jurkat cell lysate (20 μg)
  Lanes 1 - 4: Merged signal (red and green). Green - ab31828 observed at 40 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602, observed at 37 kDa.
  
  ab31828 was shown to specifically react with p38 when p38 knockout samples were used. Wild-type and p38 knockout samples were subjected to SDS-PAGE. ab31828 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.

  All lanes: Western blot - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

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  Western blot - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

  Western blot analysis of A431 cells serum starved overnight (lanes 1 & 3) or treated with pervanadate (1 mM) for 30 minutes (lanes 2 & 4). The blot was probed with anti-p38alpha (lanes 1 & 2) or anti-p38 (T180/Y182) (lanes 3-4). Lanes 5-7 shows a blot of A431 cells treated with pervanadate and probed with anti-p38 (T180/Y182) in the presence of no peptide (lane 5), phospho-ERK1 (T202/Y204) peptide (lane 6) or phoshpo-p38 (T180/Y182) peptide (lane 7).

  All lanes: Western blot - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

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  Western blot - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

  Lane 1: Wild-type HAP1 cell lysate (20 μg)
  Lane 2: p38 knockout HAP1 cell lysate (20 μg)
  Lane 3: HeLa cell lysate (20 μg)
  Lane 4: Jurkat cell lysate (20 μg)
  
  Lanes 1 - 4: Merged signal (red and green). Green - ab31828 observed at 40 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
  

  This western blot image is a comparison between ab31828 and a competitor's top cited rabbit polyclonal antibody.

  All lanes: Western blot - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

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  Immunocytochemistry/ Immunofluorescence - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

  Immunocytochemical labeling of activated p38 MAPK in pervanadate-treated mouse with ab31828. The cells were labeled with mouse monoclonal p38α MAPK and p38 MAPK antibodies, then the antibodies were detected using appropriate secondary antibodies conjugated to Cy3.

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  Flow Cytometry (Intracellular) - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

  Overlay histogram showing A431 cells stained with ab31828 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab21828, 1:100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1:500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in A431 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
  
  

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  This image is courtesy of an anonymous customer review.

  Immunocytochemistry/ Immunofluorescence - Anti-p38 delta/MAPK13 + p38 alpha/MAPK14 antibody [M138] (ab31828)

  Immunocytochemical analysis of mouse embryo fibroblast cells (NIH-3T3), labelling p38 with ab31828. Sample fixed in paraformaldehyde and blocked with 1% Donkey Serum + 1% BSA + 0.1% Triton X-100, in PBS for 30 minutes at 22°C. Incubated with ab31828 diluted 1/500 in 1% Donkey Serum + 1% BSA + 0.1% Triton X for 1 hour at 22°C. Secondary antibody was a Donkey anti-Mouse polyclonal conjugated to Alexa Fluor® 488, diluted 1/500.