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Anti-PTPN6 Antibody Y476
Also for PTPN6 (NM_002831)
|A synthetic peptide corresponding to residues in the C-term of human DHP-1 was used as immunogen. This antibody is predicted to detect isoforms 2 and 3 based on sequence analysis.|
|Mouse, Rat, Human
||Lot dependent; please refer to CoA along with shipment
|WB, IHC, IF, IP, FC
||ICC/IF: Use a concentration of 1 - 5 ug/ml; WB: 1:1000; IHC-P: Use at an assay dependent dilution; FC: 1:20; IP: 1:50
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Protein A purified
|Homo sapiens protein tyrosine phosphatase, non-receptor type 6 (PTPN6), transcript variant 1|
|HCP; HCPH; HPTP1C; PTP-1C; SH-PTP1; SHP-1; SHP-1L; SHP1|
|SHP-1 (also called PTP1C, SH-PP1, HCP, SHP and PTPN6) is a protein tyrosine phosphatase (PTP) predominantly expressed in hematopoietic cells, and regulates multiple signaling pathways involved in hematopoietic cell growth, differentiation and activation (1). After cell stimulation, SHP-1 (as well as SHP-2), translocate from cytosol to plasma membrane and bind to the tyrosine-phosphorylated receptors through their SH2 domains, becoming activated in the process (2). SHP-1 has been involved in the negative regulation of various growth promoting receptors including receptor tyrosine kinase (c-Kit, CSF-1, EGF), cytokine receptors (IL-3, IFN-a/b) and other tyrosine based receptor CD22, B & T cell Ag receptor, Lck, Zap-70 and Vav (3-5). |
EGFR1 Signaling Pathway
Jak-STAT signaling pathway
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Western blot - SHP1 antibody [Y476]; All lanes : Anti-SHP1 antibody [Y476] at 1/1000 dilution.Lane 1 : A- A431 cell lysate.Lane 2 : B- Jurkat cell lysate.Lane 3 : C- K562 cell lysate.Predicted band size : 68 kDa.Observed band size : 65 kDa .
Immunohistochemistry (Paraffin-embedded sections) - SHP1 antibody [Y476]; TA300532, at a 1/50 dilution, staining human lymph node by immunohistochemistry, Paraffin embedded tissue.
Immunocytochemistry/ Immunofluorescence-SHP1 antibody [Y476](TA300532); ICC/IF image of TA300532 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody overnight at +4°C. The secondary antibody (green) was Alexa Fluor 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43uM.
Immunoprecipitation - SHP1 antibody [Y476]; Mouse whole cell lysate prepared from splenocytes added at 10^6 cells.Immunoprecipitate step performed using Protein A/G.TA300532 used at a 1/500 dilution.For WB an HRP-rabbit anti-SHP1 (C-terminal) was used at a 1/1000 dilution
Flow Cytometry-Anti-SHP1 antibody [Y476](TA300532); Overlay histogram showing Raji cells stained with TA300532 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1ug/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.