Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM,ELISA

Reactivity: Mouse,Rat (predicted:Human,Chicken,Dog,Pig,Horse,Rabbit)

IgG

Mouse,Rat (predicted:Human,Chicken,Dog,Pig,Horse,Rabbit)

1. phosphopeptide
non phosphopeptide
2. Blank control (blue): HepG2 (fixed with 2% paraformaldehyde for 10 min at 37°C). Primary Antibody: Rabbit Anti-phospho-p38 MAPK (Thr180 + Tyr182) antibody (TMAB-01473,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.
3. Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (phospho-p38 MAPK (Thr180 + Tyr182)) Polyclonal Antibody, Unconjugated (TMAB-01473) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Tissue/cell: Hela 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 (phospho-p38 MAPK (Thr180 + Tyr182)) polyclonal Antibody, Unconjugated (TMAB-01473) 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-phospho-p38 MAPK (Thr180 + Tyr182) antibody (TMAB-01473)
Dilution: 1 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. NIH/3T3 (M) cells were treated with UV for 30 min,PC12 (R) cells were treated with UV for 30 min, 25 μg total protein per Lane of cell lysates probed with phospho-p38 MAPK (Thr180 + Tyr182) polyclonal antibody, unconjugated (TMAB-01473) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.

WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM,ELISA

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1μg /test; ELISA: 1:5000-10000

Polyclonal

KLH conjugated Synthesised phosphopeptide: human p38 MAPK around the phosphorylation site of Thr180/Tyr182

Human

1432

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