p38 alpha / MAPK14 Recombinant Rabbit Monoclonal Antibody [SR43-04] - BSA and Azide free

Western blot analysis of p38 alpha / MAPK14 on different lysates with Rabbit anti-p38 alpha / MAPK14 antibody (HA750040) at 1/1,000 dilution.

Lane 1: Hela cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: PC-12 cell lysate
Lane 4: THP-1 cell lysate
Lane 5: Jurkat cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 41 kDa
Observed band size: 38 kDa

Exposure time: 2 minutes;
12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750040) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Western blot analysis of p38 alpha / MAPK14 on different lysates with Rabbit anti-p38 alpha / MAPK14 antibody (HA750040) at 1/1,000 dilution.

Lane 1: THP-1 cell lysate (10 µg/Lane)
Lane 2: C2C12 cell lysate (10 µg/Lane)
Lane 3: PC-12 cell lysate (10 µg/Lane)
Lane 4: Rat kidney tissue lysate (20 µg/Lane)
Lane 5: Mouse kidney tissue lysate (20 µg/Lane)

Predicted band size: 41 kDa
Observed band size: 38 kDa

Exposure time: 3 minutes 10 seconds;
10% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750040) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

☑ Knockdown (KD)

Western blot analysis of p38 alpha / MAPK14 on different lysates with Rabbit anti-p38 alpha / MAPK14 antibody (HA750040) at 1/2,000 dilution.

Lane 1: A549-WT cell lysate
Lane 2: A549-KD p38 alpha / MAPK14 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 41 kDa
Observed band size: 38 kDa

Exposure time: 3 minutes; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750040) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Cell treatment (CT)

Immunocytochemistry analysis of HeLa cells treated with UV for 30 minutes then recover 30 minutes labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (HA750040) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-p38 alpha / MAPK14 antibody (HA750040) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Immunocytochemistry analysis of RAW264.7 cells labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (HA750040) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-p38 alpha / MAPK14 antibody (HA750040) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Immunocytochemistry analysis of C6 cells labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (HA750040) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-p38 alpha / MAPK14 antibody (HA750040) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of HeLa cells labeling p38 alpha / MAPK14.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA750040, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).