Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-HMGB1 antibody [EPR3506] - BSA and Azide free (ab247536)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3506] to HMGB1 - BSA and Azide free
  • Suitable for: IHC-P, WB, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-HMGB1 antibody [EPR3506] - BSA and Azide free
    See all HMGB1 primary antibodies

  • Description

    Rabbit monoclonal [EPR3506] to HMGB1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, Flow Cytmore details
    Unsuitable for: ICC or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HAP1, HeLa and Jurkat cell lysates.

  • General notes

    ab247536 is the carrier-free version of ab92310. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

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

    ab247536 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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-HMGB1 antibody [EPR3506] - BSA and Azide free (ab247536)
    Western blot - Anti-HMGB1 antibody [EPR3506] - BSA and Azide free (ab247536)
    All lanes : Anti-HMGB1 antibody [EPR3506] (ab92310) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : HMGB1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 25 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab92310).

      Lanes 1- 2: Merged signal (red and green). Green - ab92310 observed at 30 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab92310 was shown to react with HMGB1 in wild-type HeLa cells in western blot. Loss of signal at the expected size was observed when knockout cell line ab255395 (knockout cell lysate ab263782) was used. The band observed in lane 2 below 25kDa may represent truncated forms and cleaved fragments. Wild-type HeLa and HMGB1 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. ab92310 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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.

  • Western blot - Anti-HMGB1 antibody [EPR3506] - BSA and Azide free (ab247536)
    Western blot - Anti-HMGB1 antibody [EPR3506] - BSA and Azide free (ab247536)
    All lanes : Anti-HMGB1 antibody [EPR3506] (ab92310) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : HMGB1 knockout HAP1 whole cell lysate
    Lane 3 : Jurkat whole cell lysate
    Lane 4 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 25 kDa



     

    This data was developed using the same antibody clone in a different buffer formulation (ab92310).

    Lanes 1 - 4: Merged signal (red and green). Green - ab92310 observed at 30 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab92310 was shown to recognize HMGB1 in wild-type HAP1 cells as signal was lost at the expected MW in HMGB1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and HMGB1 knockout samples were subjected to SDS-PAGE. ab92310 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Anti-HMGB1 antibody [EPR3506] - BSA and Azide free (ab247536)
    Anti-HMGB1 antibody [EPR3506] - BSA and Azide free (ab247536)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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