Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-HMGB2 antibody [EPR6302] (ab133540)

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Western blot - Anti-HMGB2 antibody [EPR6302] (ab133540)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGB2 antibody [EPR6302] (ab133540)
  • Western blot - Anti-HMGB2 antibody [EPR6302] (ab133540)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGB2 antibody [EPR6302] (ab133540)
  • Western blot - Anti-HMGB2 antibody [EPR6302] (ab133540)
  • OI-RD Scanning - Anti-HMGB2 antibody [EPR6302] (ab133540)
  • Anti-HMGB2 antibody [EPR6302] (ab133540)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6302] to HMGB2
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-HMGB2 antibody [EPR6302]
    See all HMGB2 primary antibodies

  • Description

    Rabbit monoclonal [EPR6302] to HMGB2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: ICC or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide corresponding to residues in the Human HMGB2 protein (UniProt ID: P26583).

  • Positive control

    • WB: HEK-293T, HAP1, K562, HL-60, C6, RAW 264.7, and PC-12 cell lysates. IHC-P: Human breast tissue and Human colon carcinoma tissue.

  • 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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.

  • Dissociation constant (KD)

    KD = 2.97 x 10 -10 M
    Learn more about KD

  • Storage buffer

    pH: 7.2
    Preservative: 0.05% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...

  • Purity

    Tissue culture supernatant

  • Clonality

    Monoclonal

  • Clone number

    EPR6302

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133540 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 - 1/10000. Predicted molecular weight: 24 kDa.
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 24 kDa.
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for ICC or IP.

    • Target

    Images

    • Western blot - Anti-HMGB2 antibody [EPR6302] (ab133540)
      Western blot - Anti-HMGB2 antibody [EPR6302] (ab133540)
      All lanes : Anti-HMGB2 antibody [EPR6302] (ab133540) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : HMGB2 knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 24 kDa
      Observed band size: 24 kDa



      Lanes 1- 2: Merged signal (red and green). Green - ab133540 observed at 24 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab133540 was shown to react with HMGB2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266358 (knockout cell lysate ab257156) was used. Wild-type HEK-293T and HMGB2 knockout HEK-293T 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. ab133540 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGB2 antibody [EPR6302] (ab133540)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGB2 antibody [EPR6302] (ab133540)

      Immunohistochemistry analysis of paraffin embedded Human breast tissue labelling HMGB2 with ab133540 at 1/250 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-HMGB2 antibody [EPR6302] (ab133540)
      Western blot - Anti-HMGB2 antibody [EPR6302] (ab133540)
      All lanes : Anti-HMGB2 antibody [EPR6302] (ab133540) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : HMGB2 knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 24 kDa
      Observed band size: 24 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab133540 observed at 24 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab133540 was shown to recognize HMGB2 in wild-type HAP1 cells as signal was lost at the expected MW in HMGB2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and HMGB2 knockout samples were subjected to SDS-PAGE. Ab133540 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGB2 antibody [EPR6302] (ab133540)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGB2 antibody [EPR6302] (ab133540)

      Immunohistochemistry analysis of paraffin embedded Human colon carcinoma tissue labelling HMGB2 with ab133540 at 1/250 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-HMGB2 antibody [EPR6302] (ab133540)
      Western blot - Anti-HMGB2 antibody [EPR6302] (ab133540)
      All lanes : Anti-HMGB2 antibody [EPR6302] (ab133540) at 1/1000 dilution

      Lane 1 : K562 cell lysate
      Lane 2 : HL-60 cell lysate
      Lane 3 : C6 cell lysate
      Lane 4 : RAW 264.7 cell lysate
      Lane 5 : PC-12 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 24 kDa

    • OI-RD Scanning - Anti-HMGB2 antibody [EPR6302] (ab133540)
      OI-RD Scanning - Anti-HMGB2 antibody [EPR6302] (ab133540)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-HMGB2 antibody [EPR6302] (ab133540)
      Anti-HMGB2 antibody [EPR6302] (ab133540)

    References (2)

    Publishing research using ab133540? Please let us know so that we can cite the reference in this datasheet.

    ab133540 has been referenced in 2 publications.

    • Mo Y  et al. MicroRNA-329 upregulation impairs the HMGB2/ß-catenin pathway and regulates cell biological behaviors in melanoma. J Cell Physiol 234:23518-23527 (2019). PubMed: 31219186
    • Li H  et al. Centromere protein U facilitates metastasis of ovarian cancer cells by targeting high mobility group box 2 expression. Am J Cancer Res 8:835-851 (2018). PubMed: 29888106

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