Anti-Phospho-TrkA(Tyr674 + Tyr675) + TrkB(Tyr706 + Tyr707) antibody (bs-3457R)

Anti-Phospho-TrkA(Tyr674 + Tyr675) + TrkB(Tyr706 + Tyr707) antibody

DATASHEET

Host:Rabbit

Target Protein:Phospho-TrkA(Tyr674 + Tyr675) + TrkB(Tyr706 + Tyr707)

IR:Immunogen Range:TD(p-Y)(p-Y)RV

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:4914

Swiss Prot:P04629

Source:KLH conjugated Synthesised phosphopeptide derived from human TrkA around the phosphorylation site of Tyr674/675:TD(p-Y)(p-Y)RV 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:The Trk family of nerve growth factor receptors includes Trk A(also referfed to as Trk A gp140),Trk B and Trk C. The prototype member of this gene family, Trk A, encodes a 140 kDa cell surface receptor , gp140, the expression of which is restricted in vivo to neurons of the sensory spinal and cranial gangliaof neurocrest origin. Nerve growth factor (NGF) stimulates tyrosine phosphorylation of Trk gp 140 in neural cell lines and in embryonic dorsal root ganglia. By comparison, BDNF and to a lesser extent, NT-3, but not NGF, can induce tyrosine phophorylayion of Trk B gp 145. The third member of the Trk receptor family, Trk C incodes a 140 kDa protein, Trk C gp140, that is preferentially expressed in brain tissue and primarily functions as a receptor for NT-3.An additional component of the Trk receptor complex, NGFR p175, binds to neurotrophic factors with low affinity but is required for efficient signaling. NGFR p175 accelerates Trk activation and may recruit downstream dffector molecules to the ligand-bound receptor complex.

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
IHC-P(1:100-500)
IHC-F(1:100-500)
IF(1:100-500)

Cross Reactive Species:Human
Mouse
Rat
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Sample:
Lane 1: Cerebrum (Mouse) Lysate at 40 ug
Lane 2: Heart (Rat) Lysate at 40 ug
Lane 3: Large intestine (Rat) Lysate at 40 ug
Lane 4: Stomach (Rat) Lysate at 40 ug
Lane 5: SH-SY5Y (Human) Cell Lysate at 30 ug
Lane 6: K562 (Human) Cell Lysate at 30 ug
Primary: Anti-Phospho-TrkA(Tyr674 + Tyr675) + TrkB(Tyr706 + Tyr707) (bs-3457R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 140/110 kD
Observed band size: 120 kD

Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Phospho-TrkA(Tyr674/675)/TrkB(Tyr706/707) Polyclonal Antibody, Unconjugated (bs-3457R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Blank control:SH-SY5Y.
Primary Antibody (green line): Rabbit Anti-phospho-trka(tyr674 + tyr675) + trkb(tyr706 + tyr707) antibody (bs-3457R-FITC)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min 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. Acquisition of 20,000 events was performed.

Blank control:SH-SY5Y.
Primary Antibody (green line): Rabbit Anti-phospho-trka(tyr674 + tyr675) + trkb(tyr706 + tyr707) antibody (bs-3457R-FITC)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min 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. Acquisition of 20,000 events was performed.