概述

  • 产品名称
    Anti-SHP2抗体[Y478]
    参阅全部 SHP2 一抗
  • 描述
    兔单克隆抗体[Y478] to SHP2
  • 宿主
    Rabbit
  • 特异性
    ab32083 recognises SHP2. This antibody is predicted to detect splice isoform 2 based on sequence analysis.
  • 经测试应用
    适用于: WB, IHC-P, Flow Cyt, IP, ICC/IFmore details
  • 种属反应性
    与反应: Human
  • 免疫原

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

  • 阳性对照
    • IHC-P: Human breast carcinoma tissue. WB: Jurkat cell lysate. ICC/IF: Hek293 cells. Flow Cyt: HAP1-WT cells
  • 常规说明

    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 rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab32083 in the following tested applications.

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

应用 Ab评论 说明
WB 1/1000 - 1/10000. Predicted molecular weight: 68 kDa.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use a concentration of 0.1 µg/ml.
IP 1/50.
ICC/IF 1/100.

靶标

  • 功能
    Acts downstream of various receptor and cytoplasmic protein tyrosine kinases to participate in the signal transduction from the cell surface to the nucleus.
  • 组织特异性
    Widely expressed, with highest levels in heart, brain, and skeletal muscle.
  • 疾病相关
    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.
  • 序列相似性
    Belongs to the protein-tyrosine phosphatase family. Non-receptor class 2 subfamily.
    Contains 2 SH2 domains.
    Contains 1 tyrosine-protein phosphatase domain.
  • 结构域
    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.
  • 翻译后修饰
    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.
  • 细胞定位
    Cytoplasm.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

图片

  • 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 (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.

  • 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, ab8245, observed at 37 kDa.
    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 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 (ab216773) and 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.

  • 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.

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

  • Anti-SHP2 antibody [Y478] (ab32083) at 1/5000 dilution + Jurkat cell lysate

    Predicted band size: 68 kDa
    Observed band size: 70 kDa (why is the actual band size different from the predicted?)

文献

This product has been referenced in:
  • Peng YC  et al. Combination of 5-fluorouracil and 2-morphilino-8-phenyl-4H-chromen-4-one may inhibit liver cancer stem cell activity. Tumour Biol N/A:N/A (2016). Human . Read more (PubMed: 26886287) »
  • Maeshima K  et al. Abnormal PTPN11 enhancer methylation promotes rheumatoid arthritis fibroblast-like synoviocyte aggressiveness and joint inflammation. JCI Insight 1:N/A (2016). IHC ; Human . Read more (PubMed: 27275015) »

See all 5 Publications for this product

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