Anti-Phospho-TAK1 (Thr187) Polyclonal Antibody is a Rabbit antibody targeting Phospho-TAK1 (Thr187). Anti-Phospho-TAK1 (Thr187) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. Paraformaldehyde-fixed, paraffin embedded (Human stomach); Antigen retrieval by microwave in sodium citrate buffer (pH6.0); Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3% BSA) at RT for 30 min; Antibody incubation with (Phospho-TAK1 (Thr187)) Polyclonal Antibody, Unconjugated (TMAB-01515) at 1:400 overnight at 4°C, followed by conjugation to the secondary antibody (labeled with HRP) and DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by microwave in sodium citrate buffer (pH6.0); Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3% BSA) at RT for 30 min; Antibody incubation with (Phospho-TAK1 (Thr187)) Polyclonal Antibody, Unconjugated (TMAB-01515) at 1:400 overnight at 4°C, followed by conjugation to the secondary antibody (labeled with HRP) and DAB staining.
3. Sample:
Muscle (Mouse) Lysate at 40 μg
Primary: Anti-Phospho-TAK1 (Thr187) (TMAB-01515) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 67 kDa
Observed band size: 70 kDa
4. Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by microwave in sodium citrate buffer (pH6.0); Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3% BSA) at RT for 30 min; Antibody incubation with (Phospho-TAK1 (Thr187)) Polyclonal Antibody, Unconjugated (TMAB-01515) at 1:400 overnight at 4°C, followed by conjugation to the secondary antibody (labeled with HRP) and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); 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-TAK1 (Thr187)) Polyclonal Antibody, Unconjugated (TMAB-01515) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. Sample:
Lane 1: MCF-7 (Human) Cell Lysate at 30 μg
Lane 2: Hela (Human) Cell Lysate at 30 μg
Lane 3: A431 (Human) Cell Lysate at 30 μg
Lane 4: 293T (Human) Cell Lysate at 30 μg
Lane 5: K562 (Human) Cell Lysate at 30 μg
Primary:
Anti-Phospho-TAK1 (Thr187) (TMAB-01515) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 78 kDa
Observed band size: 75 kDa
7. Blank control (Black line): Raji (Black).
Primary Antibody (green line): Rabbit Anti-Phospho-TAK1 (Thr187) antibody (TMAB-01515)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 70% ice-cold methanol overnight at 4°C and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature (The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 20 min on ice.). Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.

WB,IHC-P,IHC-Fr,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human TAK1 around the phosphorylation site of Thr187

Human

Necroptosis,MAPK pathway,Inflammatory mediators,TLR Signaling,MAPK Pathway,Receptor Tyrosine Kinases,NFkB Pathway