Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-SHP2 antibody [Y478] - BSA and Azide free (ab182179)

Overview

  • Product name
    Anti-SHP2 antibody [Y478] - BSA and Azide free
    See all SHP2 primary antibodies
  • Description
    Rabbit monoclonal [Y478] to SHP2 - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, IP, IHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human SHP2 aa 500-600 (C terminal). The exact sequence is proprietary.

  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    ab182179 is a PBS-only buffer format of ab32083. Please refer to ab32083 for recommended dilutions, protocols, and image data. Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information. 

     

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab182179 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 68 kDa.

Target

  • Function
    Acts downstream of various receptor and cytoplasmic protein tyrosine kinases to participate in the signal transduction from the cell surface to the nucleus.
  • Tissue specificity
    Widely expressed, with highest levels in heart, brain, and skeletal muscle.
  • Involvement in disease
    Defects in PTPN11 are the cause of LEOPARD syndrome type 1 (LEOPARD1) [MIM:151100]. It is an autosomal dominant disorder allelic with Noonan syndrome. The acronym LEOPARD stands for lentigines, electrocardiographic conduction abnormalities, ocular hypertelorism, pulmonic stenosis, abnormalities of genitalia, retardation of growth, and deafness.
    Defects in PTPN11 are the cause of Noonan syndrome type 1 (NS1) [MIM:163950]. Noonan syndrome (NS) is a disorder characterized by dysmorphic facial features, short stature, hypertelorism, cardiac anomalies, deafness, motor delay, and a bleeding diathesis. Some patients with Noonan syndrome type 1 develop multiple giant cell lesions of the jaw or other bony or soft tissues, which are classified as pigmented villomoduolar synovitis (PVNS) when occurring in the jaw or joints. Note=Mutations in PTPN11 account for more than 50% of the cases. Rarely, NS is associated with juvenile myelomonocytic leukemia (JMML). NS1 inheritance is autosomal dominant.
    Defects in PTPN11 are a cause of juvenile myelomonocytic leukemia (JMML) [MIM:607785]. JMML is a pediatric myelodysplastic syndrome that constitutes approximately 30% of childhood cases of myelodysplastic syndrome (MDS) and 2% of leukemia. It is characterized by leukocytosis with tissue infiltration and in vitro hypersensitivity of myeloid progenitors to granulocyte-macrophage colony stimulating factor.
    Defects in PTPN11 are a cause of metachondromatosis (MC) [MIM:156250]. It is a skeletal disorder with radiologic fetarures of both multiple exostoses and Ollier disease, characterized by the presence of multiple enchondromas and osteochondroma-like lesions.
  • Sequence similarities
    Belongs to the protein-tyrosine phosphatase family. Non-receptor class 2 subfamily.
    Contains 2 SH2 domains.
    Contains 1 tyrosine-protein phosphatase domain.
  • Domain
    The SH2 domains repress phosphatase activity. Binding of these domains to phosphotyrosine-containing proteins relieves this auto-inhibition, possibly by inducing a conformational change in the enzyme.
  • Post-translational
    modifications
    Phosphorylated on Tyr-546 and Tyr-584 upon receptor protein tyrosine kinase activation; which creates a binding site for GRB2 and other SH2-containing proteins.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • BPTP3 antibody
    • CFC antibody
    • JMML antibody
    • METCDS antibody
    • MGC14433 antibody
    • NS1 antibody
    • OTTHUMP00000166107 antibody
    • OTTHUMP00000166108 antibody
    • Protein tyrosine phosphatase 2 antibody
    • Protein tyrosine phosphatase 2C antibody
    • Protein tyrosine phosphatase non receptor type 11 antibody
    • Protein-tyrosine phosphatase 1D antibody
    • Protein-tyrosine phosphatase 2C antibody
    • PTN11_HUMAN antibody
    • PTP-1D antibody
    • PTP-2C antibody
    • PTP1D antibody
    • PTP2C antibody
    • PTPN11 antibody
    • SAP2 antibody
    • SH-PTP2 antibody
    • SH-PTP3 antibody
    • SH2 domain containing protein tyrosine phosphatase 2 antibody
    • SHP 2 antibody
    • SHP-2 antibody
    • Shp2 antibody
    • SHPTP2 antibody
    • SHPTP3 antibody
    • Syp antibody
    • Tyrosine-protein phosphatase non-receptor type 11 antibody
    see all

Images

  • 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 (ab172730) instead of ab32083 in THP-1 whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32083).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor®594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 (ab150081) at 1/2000 dilution for 30 min at 22°C.

    A rabbit IgG isotype control antibody  (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32083).

  • 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)(ab150077)(1:2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32083).

  • Immunohistochemical analysis of SHP2 expression in paraffin embedded human breast carcinoma, using 1/50 unpurified ab32083.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32083).

References

ab182179 has not yet been referenced specifically in any publications.

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