Anti-p38 alpha/MAPK14 Antibody Picoband®

Western blot analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody (A00176-2).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: human SW620 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human CACO-2 whole cell lysates,
Lane 5: rat lung tissue lysates,
Lane 6: rat heart tissue lysates,
Lane 7: mouse lung tissue lysates,
Lane 8: mouse heart tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-p38 alpha/MAPK14 antigen affinity purified polyclonal antibody (A00176-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for p38 alpha/MAPK14 at approximately 38 kDa. The expected band size for p38 alpha/MAPK14 is at 41 kDa.

Western blot analysis of p38 alpha using anti-p38 alpha antibody (A00176-2).
Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 μg of sample under reducing conditions.
Lane 1: mouse spleen tissue lysates,
Lane 2: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-p38 alpha antigen affinity purified polyclonal antibody (A00176-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween-20 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for p38 alpha at approximately 41 kDa. The expected band size for p38 alpha is at 41 kDa.

IHC analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody (A00176-2).
p38 alpha/MAPK14 was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-p38 alpha/MAPK14 Antibody (A00176-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody (A00176-2).
p38 alpha/MAPK14 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-p38 alpha/MAPK14 Antibody (A00176-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody (A00176-2).
p38 alpha/MAPK14 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-p38 alpha/MAPK14 Antibody (A00176-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of P38 Alpha/MAPK14 using anti-P38 Alpha/MAPK14 antibody (A00176-2) and anti-Alpha Tubulin antibody (M03989-3).
P38 Alpha/MAPK14 was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-P38 Alpha/MAPK14 Antibody (A00176-2) and mouse anti-Alpha Tubulin antibody (M03989-3) overnight at 4°C. Fluoro488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunoprecipitating p38 alpha/MAPK14 in Jurkat whole cell lysate.
Western blot analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody (A00176-2);
Lane 1: Jurkat whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-p38 alpha/MAPK14 antibody in Jurkat whole cell lysate;
Lane 3: anti-p38 alpha/MAPK14 antibody (2μg) + Jurkat whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-p38 alpha/MAPK14 antigen affinity purified polyclonal antibody (A00176-2) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for p38 alpha/MAPK14 at approximately 38 kDa. The expected band size for p38 alpha/MAPK14 is at 41 kDa.

Flow Cytometry analysis of CACO-2 cells using anti-P38 Alpha/MAPK14 antibody (A00176-2).
Overlay histogram showing CACO-2 cells stained with A00176-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-P38 Alpha/MAPK14 Antibody (A00176-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

(A) Protein expression levels of LC3-I, LC3-II, p38 MAPK, and p-p38 MAPK in SMMC-7721 cells measured by Western blot after treatment with 5 μM PL, 10 μM 3-MA autophagy inhibitor, 10 μM rapamycin autophagy agonist, 10 μM SB202190, and 10 μM SB203580 p38 MAPK inhibitor. (B) LC3-I, LC3-II, p38 MAPK, and p-p38 MAPK were used as internal reference total proteins for gray value analysis. * p < 0.05, ** p < 0.01 *** p < 0.001, compared with the PL group. # p < 0.05, ## p < 0.01 ### p < 0.001 vs. the control group. (C) Protein expression levels of LC3-I, LC3-II, p38 MAPK, and p-p38 MAPK in BEL-7404 cells measured by Western blot after treatment with 5 μM PL, 10 μM 3-MA autophagy inhibitor, 10 μM rapamycin autophagy agonist, 10 μM SB202190, and 10 μM SB203580 p38 MAPK inhibitor. (D) LC3-I, LC3-II, p38 MAPK, and p-p38 MAPK were used as internal reference total proteins for gray value analysis. * p < 0.05, ** p < 0.01 *** p < 0.001, compared with the PL group. # p < 0.05, ## p < 0.01 ### p < 0.001 vs. the control group.
Index in PubMed under a CC BY license. PMID: 33912033