Rabbit anti-Human, Mouse SHP1/PTPN6 Polyclonal Antibody | anti-PTPN6 antibody
Anti-SHP1/PTPN6 Antibody
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of SHP1/PTPN6 using anti- SHP1/PTPN6 antibody (MBS1753384).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Jurkat whole cell lysatesLane 2: human Raji whole cell lysatesLane 3: mouse Raw264. 7 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti- SHP1/PTPN6 antigen affinity purified polyclonal antibody (Catalog # MBS1753384) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for SHP1/PTPN6 at approximately 68KD. The expected band size for SHP1/PTPN6 is at 68KD. )
Immunofluorescence (IF)
(Figure 2. IF analysis of SHP1/PTPN6 using anti-SHP1/PTPN6 antibody (MBS1753384).SHP1/PTPN6 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-SHP1/PTPN6 Antibody (MBS1753384) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of A431 cells using anti-SHP1/PTPN6 antibody (MBS1753384).Overlay histogram showing A431 cells stained with MBS1753384 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SHP1/PTPN6 Antibody (MBS1753384, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Beghini, A., Ripamonti, C. B., Peterlongo, P., Roversi, G., Cairoli, R., Morra, E., Larizza, L. RNA hyperediting and alternative splicing of hematopoietic cell phosphatase (PTPN6) gene in acute myeloid leukemia. Hum. Molec. Genet. 9: 2297-2304, 2000.
3. Croker, B. A., Lawson, B. R., Rutschmann, S., Berger, M., Eidenschenk, C., Blasius, A. L., Moresco, E. M. Y., Sovath, S., Cengia, L., Shultz, L. D., Theofilopoulos, A. N., Pettersson, S., Beutler, B. A. Inflammation and autoimmunity caused by a SHP1 mutation depend on IL-1, MyD88, and microbial trigger. Proc. Nat. Acad. Sci. 105: 15028-15033, 2008. Note: Erratum: Proc. Nat. Acad. Sci. 105: 19561 only, 2008.
NCBI and Uniprot Product Information
NCBI Description
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]
Uniprot Description
Function: Modulates signaling by tyrosine phosphorylated cell surface receptors such as KIT and the EGF receptor/EGFR. The SH2 regions may interact with other cellular components to modulate its own phosphatase activity against interacting substrates. Together with MTUS1, induces UBE2V2 expression upon angiotensin II stimulation. Plays a key role in hematopoiesis. Ref.20
Catalytic activity: Protein tyrosine phosphate + H2O = protein tyrosine + phosphate.
Subunit structure: Monomer. Interacts with MTUS1
By similarity. Interacts with MILR1 (tyrosine-phosphorylated). Interacts with KIT
By similarity. Binds PTPNS1, LILRB1 and LILRB2. Interacts with FCRL2, FCRL3, FCRL4, CD300LF, CDK2 and CD84. Interacts with KIR2DL1; the interaction is enhanced by ARRB2. Interacts (via SH2 1 domain) with ROS1; the interaction is direct and promotes ROS1 dephosphorylation. Interacts with EGFR; inhibits EGFR-dependent activation of MAPK/ERK. Interacts with LYN. Interacts with the tyrosine phosphorylated form of PDPK1. Ref.14 Ref.15 Ref.16 Ref.17 Ref.18 Ref.19 Ref.20 Ref.21 Ref.22 Ref.25 Ref.26 Ref.29
Subcellular location: Cytoplasm. Nucleus. Note: In neurons, translocates into the nucleus after treatment with angiotensin II
By similarity. Shuttles between the cytoplasm and nucleus via its association with PDPK1. Ref.26 Ref.29
Tissue specificity: Isoform 1 is expressed in hematopoietic cells. Isoform 2 is expressed in non-hematopoietic cells.
Domain: The N-terminal SH2 domain functions as an auto-inhibitory domain, blocking the catalytic domain in the ligand-free close conformation. Ref.31 Ref.33
Post-translational modification: Phosphorylated on tyrosine residues. Binding of KITLG/SCF to KIT increases tyrosine phosphorylation
By similarity. Phosphorylation at Tyr-564 enhances phosphatase activity. Ref.13 Ref.17
Sequence similarities: Belongs to the protein-tyrosine phosphatase family. Non-receptor class 2 subfamily.Contains 2 SH2 domains.Contains 1 tyrosine-protein phosphatase domain.
