Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22468-6] to PTP1B - BSA and Azide free
  • Suitable for: WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free
    See all PTP1B primary antibodies

  • Description

    Rabbit monoclonal [EPR22468-6] to PTP1B - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IPmore details
    Unsuitable for: Flow Cyt or ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HCT116, HeLa, HepG2, A549, MCF7, SW480, RAW264.7, NIH/3T3 and TF-1 whole cell lysate. Wild-type HAP1 whole cell lysate. Human placental cell lysate. Human fetal kidney cell lysate. Mouse brain and spleen cell lysate. Rat brain and heart cell lysate. IHC-P: Human breast cancer and gastric cancer tissue. Mouse and rat spleen tissue. HCT116 whole cell lysate.

  • General notes

    ab254269 is the carrier-free version of ab252928.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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 production
    For more information see here.

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

Properties

Images

  • Western blot - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)
    Western blot - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)
    All lanes : Anti-PTP1B antibody [EPR22468-6] (ab252928) at 1/2000 dilution

    Lane 1 : HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate
    Lane 2 : Wild-type HAP1 whole cell lysate
    Lane 3 : PTP1B knockout HAP1 whole cell lysate
    Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 5 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
    Lane 6 : TF-1 (human Erythroleukemia erythroblast), whole cell lysate
    Lane 7 : A549 (human lung 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
    Observed band size: 36,46,48,50 kDa
    why is the actual band size different from the predicted?



    ab252928 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. ab252928 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/2000 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 50kDa may represent truncated forms and cleaved fragments.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lanes 1-3: 3 mins; Lanes 4-6: 15 secs; Lane 7: 3 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252928).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)

    Immunohistochemical analysis of paraffin-embedded human breast cancer (panel A) and its adjacent non-cancerous tissue (panel B) labeling PTP1B with ab252928 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Higher cytoplasmic staining in human breast cancer tissue (panel A) while weakly staining in its adjacent non-cancerous tissue (panel B) (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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252928).
  • Immunoprecipitation - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)
    Immunoprecipitation - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)

    PTP1B was immunoprecipitated from 0.35 mg HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate with ab252928 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab252928 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

    Lane 1: HCT116 whole cell lysate 10 µg (Input).
    Lane 2: ab252928 IP in HCT116 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252928 in HCT116 whole cell lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 3 seconds.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252928).

  • Western blot - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)
    Western blot - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)
    All lanes : Anti-PTP1B antibody [EPR22468-6] (ab252928) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : PTPN1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 50 kDa
    Observed band size: 50 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab252928).

      Lanes 1- 2: Merged signal (red and green). Green - ab252928 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab252928 was shown to react with PTP1B in wild-type HeLa cells in western blot. The band observed in knockout cell line ab265014 (knockout cell lysate ab257617) lane below 50kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and PTPN1 knockout HeLa 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. ab252928 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated 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 (ab216773) and 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)

    Immunohistochemical analysis of paraffin-embedded human gastric cancer (panel A) and its adjacent non-cancerous tissue (panel B) labeling PTP1B with ab252928 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Higher cytoplasmic staining in human gastric cancer tissue (panel A) while weakly staining in its adjacent non-cancerous tissue (panel B) (PMID: 21442314) 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252928).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling PTP1B with ab252928 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in rat spleen (PMID:18716132) 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252928).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling PTP1B with ab252928 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in mouse spleen (PMID:18716132) 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252928).
  • Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)
    Anti-PTP1B antibody [EPR22468-6] - BSA and Azide free (ab254269)

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