Anti-Phospho-TIRAP (Tyr86) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-TIRAP (Tyr86). Anti-Phospho-TIRAP (Tyr86) Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Rat (predicted:Mouse,Dog,Pig,Cow,Horse,GuineaPig)

1. Blank control: U937.
Primary Antibody (green line): Rabbit Anti-phospho-TIRAP(Tyr86) antibody (TMAB-11381)
Dilution: 2 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-PE
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. 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. Acquisition of 20,000 events was performed.
2. Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Incubation with (phospho-TIRAP (Tyr86) ) Polyclonal Antibody, Unconjugated (TMAB-11381) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Incubation with (phospho-TIRAP (Tyr86) ) Polyclonal Antibody, Unconjugated (TMAB-11381) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (human heart); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Incubation with (phospho-TIRAP (Tyr86) ) Polyclonal Antibody, Unconjugated (TMAB-11381) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
5. Sample:
Lane 1: Human HepG2 cell lysates
Lane 2: Human HUVEC cell lysates
Lane 3: Human U251 cell lysates
Lane 4: Human SH-SY5Y cell lysates
Lane 5: Human K562 cell lysates
Primary: Anti-phospho-TIRAP (Tyr86) (TMAB-11381) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 24 kDa
Observed band size: 35 kDa

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 2μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human TIRAP around the acetylation site of Tyr86

Human

TIRAP

Toll/interleukin-1 receptor domain-containing adapter protein

Adapter involved in TLR2 and TLR4 signaling pathways in the innate immune response. Acts via IRAK2 and TRAF-6, leading to the activation of NF-kappa-B, MAPK1, MAPK3 and JNK, and resulting in cytokine secretion and the inflammatory response. Positively regulates the production of TNF-alpha and interleukin-6.