Overview

  • Product name
    Anti-PTP1B antibody [EPR22474]
    See all PTP1B primary antibodies
  • Description
    Rabbit monoclonal [EPR22474] to PTP1B
  • Host species
    Rabbit
  • Specificity

    IHC is recommended for human only.

  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human PTP1B aa 1-300. The exact sequence is proprietary.
    Database link: P18031

  • Positive control
    • WB: Wild type HAP1 whole cell lysate. HeLa, HepG2, A549, MCF7, SW480, RAW 264.7, PC-12, NIH/3T3 and HCT 116 whole cell lysate. IHC-P: Human breast cancer and colon cancer tissue. ICC/IF: HeLa, wild type HAP1 and HCT 116 cells. Flow Cyt: HeLa and HCT 116 cells. IP: PTP1B IP in HCT 116 whole cell lysate.
  • General notes

    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 ab244207 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
WB 1/1000. Predicted molecular weight: 50 kDa.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC is recommended for human only.

ICC/IF 1/50.
IP 1/30.
Flow Cyt 1/50.

Target

  • Function
    May play an important role in CKII- and p60c-src-induced signal transduction cascades.
  • Sequence similarities
    Belongs to the protein-tyrosine phosphatase family. Non-receptor class 1 subfamily.
    Contains 1 tyrosine-protein phosphatase domain.
  • Post-translational
    modifications
    Oxidized on Cys-215; the Cys-SOH formed in response to redox signaling reacts with the alpha-amido of the following residue to form a 4-amino-3-isothiazolidinone serine cross-link, triggering a conformational change that inhibits substrate binding and activity. The active site can be restored by reduction.
  • Cellular localization
    Endoplasmic reticulum membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • PTP1B antibody
    • Non receptor tyrosine phosphatase 1 antibody
    • Protein phosphotyrosylphosphatase 1B antibody
    • Protein tyrosine phosphatase 1B antibody
    • Protein tyrosine phosphatase non receptor type 1 antibody
    • Protein tyrosine phosphatase placental antibody
    • Protein-tyrosine phosphatase 1B antibody
    • PTN1_HUMAN antibody
    • PTP 1B antibody
    • PTP-1B antibody
    • PTPN 1 antibody
    • PTPN1 antibody
    • Tyrosine protein phosphatase non receptor type 1 antibody
    • Tyrosine-protein phosphatase non-receptor type 1 antibody
    see all

Images

  • All lanes : Anti-PTP1B antibody [EPR22474] (ab244207) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : PTP1B knockout HAP1 whole cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 50 kDa



    ab244207 was shown to specifically react with PTP1B in wild-type HAP1 cells as signal was lost in PTP1B knockout cells. Wild-type and PTP1B knockout samples were subjected to SDS-PAGE. ab244207 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

    The expression profile observed is consistent with what has been described in the literature (PMID: 18253097; PMID: 11895943; PMID: 19797268). The bands below 50 kDa may represent truncated forms and cleaved fragments.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • All lanes : Anti-PTP1B antibody [EPR22474] (ab244207) at 1/1000 dilution

    Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
    Lane 2 : A549 (human lung carcinoma epithelial cell) whole cell lysate
    Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 50 kDa



    The expression profile observed is consistent with what has been described in the literature (PMID: 18253097; PMID: 11895943; PMID: 19797268). The bands below 50 kDa may represent truncated forms and cleaved fragments.

    Blocking/Dilution buffer: NFDM/TBST.

    Exposure time: 37 seconds.

  • All lanes : Anti-PTP1B antibody [EPR22474] (ab244207) at 1/1000 dilution

    Lane 1 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 50 kDa



    The expression profile observed is consistent with what has been described in the literature (PMID: 18253097; PMID: 11895943; PMID: 19797268). The bands below 50 kDa may represent truncated forms and cleaved fragments.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 48 seconds.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling PTP1B with ab244207 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Cytoplasmic staining in human colon cancer (PMID:27752061) is observed. Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling PTP1B with ab244207 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Higher cytoplasmic expression in human breast cancer than that of adjacent normal tissues (PMID: 27465552) is observed. Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling PTP1B with ab244207 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in HeLa cells is observed. The nuclear counterstain is DAPI (blue).
    Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
    The negative control is the secondary antibody only.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cells labeling PTP1B with ab244207 at 1/50 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in HCT 116 cells. The nuclear counterstain is DAPI (blue).
    Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
    The negative control is the secondary antibody only.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Wild type and PTP1B-knockout HAP1 (Human chronic myelogenous leukemia near-haploid cell line) cells labeling PTP1B with ab244207 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing no staining in PTP1B-knockout HAP1 cells. The nuclear counterstain is DAPI (blue).
    Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
    The negative control is the secondary antibody only.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling PTP1B with ab244207 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730)isotype control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cell line labeling PTP1B with ab244207 at 1/50 (red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

  • PTP1B  was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate with ab244207 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab244207 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
    Lane 1: HCT 116 whole cell lysate 10 µg (Input).
    Lane 2: ab244207 IP in HCT 116 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab244207 in HCT 116 whole cell lysate.
    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 30 seconds.

    The observed MW is consistent with what described in the literatures. (PMID: 18253097; PMID: 11895943; PMID: 19797268).

References

ab244207 has not yet been referenced specifically in any publications.

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