Anti-MAPK14 Polyclonal Antibody is a Rabbit antibody targeting MAPK14. Anti-MAPK14 Polyclonal Antibody can be used in ICC/IF,FCM.

IgG

Human,Mouse (predicted:Rat,Dog,Rabbit,Sheep)

1. Blank control: HepG2 (blue). Primary Antibody: Rabbit Anti-P38 MAPK antibody (TMAB-01102,Green); Dilution: 1 μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit Igg (orange),used under the same conditions; Secondary Antibody: Goat anti-rabbit IgG-FITC (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde for 10 min at 37°C. Primary antibody (TMAB-01102, 1 μg/1x10^6 cells) were incubated for 30 min at room temperature, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 40 min at room temperature.
2. Blank control: MCF7. Primary Antibody (green line): Rabbit Anti-P38 MAPK antibody (TMAB-01102)
Dilution: 2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. Tissue/cell: HUVEC cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (P38 MAPK) polyclonal Antibody, Unconjugated (TMAB-01102) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
4. Tissue/cell: MCF7 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (P38 MAPK) polyclonal Antibody, Unconjugated (TMAB-01102) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
5. Blank control: Raw264.7. Primary Antibody (green line): Rabbit Anti-P38 MAPK antibody (TMAB-01102)
Dilution: 2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
6. The Hela (H) （UV stimulated) cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C,the cells then were incubated in 5% BSA to block non-specific protein-protein interactions (30 min at room temperature). Primary Antibody (green): Rabbit Anti-P38 MAPK/MAPK14 antibody (TMAB-01102): 1 μg/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC: 1 μg/test. Isotype Control (orange): Rabbit IgG. Blank control (black): PBS.
7. 4% Paraformaldehyde-fixed Hela (H) （UV stimulated) cell; Triton X-100 at RT for 20 min; Antibody incubation with (P38 MAPK/MAPK14) polyclonal Antibody, unconjugated (TMAB-01102) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green ) at 37°C for 90 min, DAPI (blue) was used to stain the cell nucleus. PBS instead of the primary antibody was used as the blank control.

ICC/IF,FCM

ICC/IF: 1:50-1:200; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human P38MAPK

Human

MAPK pathway,TLR Signaling,Other Cell Signaling Kits,MAPK Pathway