Anti-phospho-PTPN6 (Ser591) antibody (bs-5578R)

Anti-phospho-PTPN6 (Ser591) antibody

DATASHEET

Host:Rabbit

Target Protein:phospho-PTPN6 (Ser591)

IR:Immunogen Range:KG(p-S)LK

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:5777

Swiss Prot:P29350

Source:KLH conjugated Synthesised phosphopeptide derived from human PTPN6 around the phosphorylation site of Ser591:KG(p-S)LK 

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 protein encoded by this gene is a member of the protein tyrosine phosphatase (PTP) family. PTPs are known to be signaling molecules that regulate a variety of cellular processes including cell growth, differentiation, mitotic cycle, and oncogenic transformation. N-terminal part of this PTP contains two tandem Src homolog (SH2) domains, which act as protein phospho-tyrosine binding domains, and mediate the interaction of this PTP with its substrates. This PTP is expressed primarily in hematopoietic cells, and functions as an important regulator of multiple signaling pathways in hematopoietic cells. This PTP has been shown to interact with, and dephosphorylate a wide spectrum of phospho-proteins involved in hematopoietic cell signaling. Multiple alternatively spliced variants of this gene, which encode distinct isoforms, have been reported. [provided by RefSeq, Jul 2008]

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(1μg/Test)
IF(1:100-500)

Cross Reactive Species:Human
Mouse
Rat
Dog
Pig
Horse
.

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

VALIDATION IMAGES

Sample:
A549 Cell (Human) Lysate at 40 ug
MCF-7 Cell (Human) Lysate at 40 ug
Primary: Anti-phospho-PTPN6(Ser591) (bs-5578R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 65 kD
Observed band size: 65 kD

Paraformaldehyde-fixed, paraffin embedded (rat spleen tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PTPN6(Ser591)) Polyclonal Antibody, Unconjugated (bs-5578R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control: Hela cells(fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody:Rabbit Anti-phospho-PTPN6(Ser591) antibody(bs-5578R), Dilution: 1μg 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-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.

Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-PTPN6 antibody (bs-5578R-AF647)
Dilution: 1μg /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 90% ice-cold methanol for 20 min at -20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature.Acquisition of 20,000 events was performed.