Name: Anti-SHP2 antibody [Y478]
SKU: ab32083
Rating: 4 (1 reviews)

Rabbit Recombinant Monoclonal SHP2 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 14 publications.

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Images

Western blot - Anti-SHP2 antibody [Y478] (AB32083), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/40	Notes For unpurified use at 1/50 Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000 - 1/10000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes For unpurified use at 1/100.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes For unpurified use at 0.1 μg/ml

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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11 products for Alternative Product

Target data

See full target information PTPN11

Function

Acts downstream of various receptor and cytoplasmic protein tyrosine kinases to participate in the signal transduction from the cell surface to the nucleus (PubMed:10655584, PubMed:14739280, PubMed:18559669, PubMed:18829466, PubMed:26742426, PubMed:28074573). Positively regulates MAPK signal transduction pathway (PubMed:28074573). Dephosphorylates GAB1, ARHGAP35 and EGFR (PubMed:28074573). Dephosphorylates ROCK2 at 'Tyr-722' resulting in stimulation of its RhoA binding activity (PubMed:18559669). Dephosphorylates CDC73 (PubMed:26742426). Dephosphorylates SOX9 on tyrosine residues, leading to inactivate SOX9 and promote ossification (By similarity). Dephosphorylates tyrosine-phosphorylated NEDD9/CAS-L (PubMed:19275884).

Alternative names

PTP2C, SHPTP2, PTPN11, Tyrosine-protein phosphatase non-receptor type 11, Protein-tyrosine phosphatase 1D, Protein-tyrosine phosphatase 2C, SH-PTP2, SH-PTP3, PTP-1D, PTP-2C, SHP-2, Shp2

Recommended products

Rabbit Recombinant Monoclonal SHP2 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 14 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: Y478
Purification technique: Affinity purification Protein A
Specificity: This antibody recognises SHP2. This antibody is predicted to detect splice isoform 2 based on sequence analysis.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

SHP2 also known as PTPN11 is a protein tyrosine phosphatase with a molecular mass of approximately 68 kDa. It is expressed in various tissues including the heart liver and immune cells. SHP2 belongs to the non-receptor class of protein tyrosine phosphatases and plays a critical role in cell signaling by acting as a regulator of signal transduction processes. SHP2 mediates these processes by dephosphorylating specific phosphotyrosine residues on target proteins influencing various cellular functions like proliferation differentiation and survival.

Biological function summary

The role of SHP2 extends to involvement in several signaling cascades such as the Ras/MAPK and PI3K/AKT pathways. It functions as an essential component within protein complexes that facilitate cell communication and response to external signals. The protein modulates growth factor signaling and cytokine signaling highlighting its significance in normal cell function and development. SHP2's statement in signaling processes makes it an important regulator of cellular dynamics.

Pathways

SHP2 participates in the Ras/MAPK and PI3K/AKT signaling pathways which are important for regulating cell growth survival and differentiation. Within these pathways SHP2 interacts with various signaling molecules including Grb2 Sos and Gab family adaptors. These interactions coordinate cellular responses to growth factors and other extracellular cues ensuring proper pathway activation and control. By serving as a critical mediator SHP2 integrates signals that are necessary for appropriate cellular outcomes.

Associated diseases and disorders

SHP2 is associated with several conditions such as Noonan syndrome and various cancers. Mutations in the PTPN11 gene which encodes SHP2 often result in aberrant signaling that leads to developmental anomalies or tumorigenesis. In Noonan syndrome the mutated SHP2 protein results in disrupted Ras/MAPK pathway signaling. As for cancers SHP2 is often found to be overactive leading to enhanced cell proliferation and survival. In these contexts SHP2 is interconnected with other proteins like RAS and RAF which also contribute to oncogenic pathway activation and disease progression.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

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12 product images

Western blot - Anti-SHP2 antibody [Y478] (ab32083)
Lanes 1- 2: Merged signal (red and green). Green - ab32083 observed at 68 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
ab32083 was shown to react with SHP2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human PTPN11 (SHP2) knockout HEK-293T cell line ab266450 (knockout cell lysate Human PTPN11 (SHP2) knockout HEK-293T cell lysate ab257618) was used. Wild-type HEK-293T and PTPN11 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32083 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-SHP2 antibody [Y478] (ab32083) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: PTPN11 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (Human PTPN11 (SHP2) knockout HEK-293T cell line ab266450)
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 68 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP2 antibody [Y478] (ab32083)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human endometrium cancer tissue sections labeling SHP2 with Purified ab32083 at 1:100 dilution (5.51 µg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain
Immunoprecipitation - Anti-SHP2 antibody [Y478] (ab32083)
ab32083 (purified) at 1:40 dilution (2μg) immunoprecipitating SHP2 in THP-1 whole cell lysate.
Lane 1 (input): THP-1 (Human monocytic leukemia monocyte) whole cell lysate 10μg
Lane 2 (+): ab32083 & THP-1 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab32083 in THP-1 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-SHP2 antibody [Y478] (ab32083)
Predicted band size: 68 kDa
Flow Cytometry (Intracellular) - Anti-SHP2 antibody [Y478] (ab32083)
Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling SHP2 with purified ab32083 at 1/50 dilution (10 μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Western blot - Anti-SHP2 antibody [Y478] (ab32083)
Lane 1: Wild-type HAP1 cell lysate (20 μg)
Lane 2: SHP2 knockout HAP1 cell lysate (20 μg)
Lane 3: A431 cell lysate (20 μg)
Lane 4: Jurkat cell lysate (20 μg)
Lanes 1 to 4: Merged signal (red and green). Green - ab32083 observed at 68 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
unpurified ab32083 was shown to specifically react with SHP2 when SHP2 knockout samples were used. Wild-type and SHP2 knockout samples were subjected to SDS-PAGE. ab32083 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were both diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-SHP2 antibody [Y478] (ab32083)
Predicted band size: 68 kDa
Western blot - Anti-SHP2 antibody [Y478] (ab32083)
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-SHP2 antibody [Y478] (ab32083) at 0.3 µg/mL
All lanes: THP-1 (Human monocytic leukemia monocyte) whole cell lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 68 kDa
Immunocytochemistry/ Immunofluorescence - Anti-SHP2 antibody [Y478] (ab32083)
Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling SHP2 with Purified ab32083 at 1:50 dilution (11 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor®594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor®488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow Cytometry (Intracellular) - Anti-SHP2 antibody [Y478] (ab32083)
Overlay histogram showing HAP1 wildtype (green line) and HAP1-PTPN11 knockout cells (red line) stained with ab32083. The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (unpurified ab32083, 0.1μg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) presorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG isotype control antibody (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-PTPN11 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min) permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
Flow Cytometry (Intracellular) - Anti-SHP2 antibody [Y478] (ab32083)
Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling SHP2 with unpurified ab32083 at 1/500 dilution (1ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
Immunocytochemistry/ Immunofluorescence - Anti-SHP2 antibody [Y478] (ab32083)
unpurified ab32083 stained Hek293 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32083 at 1/100 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43μM for 1hour at room temperature.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP2 antibody [Y478] (ab32083)
Immunohistochemical analysis of SHP2 expression in paraffin embedded human breast carcinoma, using 1/50 unpurified ab32083.
Western blot - Anti-SHP2 antibody [Y478] (ab32083)
All lanes: Western blot - Anti-SHP2 antibody [Y478] (ab32083) at 1/5000 dilution
All lanes: Jurkat cell lysate
Predicted band size: 68 kDa
Observed band size: 70 kDa