Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22365-230] to Heparanase 1
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Heparanase 1 antibody [EPR22365-230]
    See all Heparanase 1 primary antibodies

  • Description

    Rabbit monoclonal [EPR22365-230] to Heparanase 1

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa and Capan-1 whole cell lysates; Recombinant human active heparinase 1 protein (aa36-109 & 158-543).

  • General notes

    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-Heparanase 1 antibody [EPR22365-230] (ab254254)
    Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)
    All lanes : Anti-Heparanase 1 antibody [EPR22365-230] (ab254254) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : HPSE knockout HeLa cell lysate
    Lane 3 : Capan-1 cell lysate
    Lane 4 : Human pancreas tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 61 kDa
    Observed band size: 52,65 kDa
    why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab254254 observed at 52,65 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab254254 Anti-Heparanase 1 antibody [EPR22365-230] was shown to specifically react with Heparanase 1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265413 (knockout cell lysate ab258456) was used. Wild-type and Heparanase 1 knockout samples were subjected to SDS-PAGE. ab254254 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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.

  • Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)
    Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)
    Anti-Heparanase 1 antibody [EPR22365-230] (ab254254) at 1/1000 dilution + Recombinant human active heparinase 1 protein (aa36-109 & 158-543, expressed in CHo cells), 20 ng

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

    Developed using the ECL technique.

    Predicted band size: 61 kDa


    Exposure time: 6 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

    This blot was developed using a higher sensitivity ECL substrate.

  • Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)
    Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)
    All lanes : Anti-Heparanase 1 antibody [EPR22365-230] (ab254254) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Capan-1 (human pancreas adenocarcinoma epithelial cell line) whole cell lysate
    Lane 3 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
    Lane 4 : Human pancreas 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: 61 kDa
    Observed band size: 50,65 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure times.

    Lane 1: 3 minutes; Lanes 2 -4: 78 seconds.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 16458254, 11406531).

    Negative control: Human pancreas, MCF7 (PMID: 11406531, 10395325).

    This blot was developed using a higher sensitivity ECL substrate.

     

  • Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)
    Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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