Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-HMGB2 antibody [EPR6301] (ab124670)

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Western blot - Anti-HMGB2 antibody [EPR6301] (ab124670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGB2 antibody [EPR6301] (ab124670)
  • Immunocytochemistry/ Immunofluorescence - Anti-HMGB2 antibody [EPR6301] (ab124670)
  • Western blot - Anti-HMGB2 antibody [EPR6301] (ab124670)
  • Western blot - Anti-HMGB2 antibody [EPR6301] (ab124670)
  • OI-RD Scanning - Anti-HMGB2 antibody [EPR6301] (ab124670)
  • Anti-HMGB2 antibody [EPR6301] (ab124670)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6301] to HMGB2
  • Suitable for: ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-HMGB2 antibody [EPR6301]
    See all HMGB2 primary antibodies

  • Description

    Rabbit monoclonal [EPR6301] to HMGB2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human HMGB2 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HEK-293T, HAP1, K562, HeLa and PC12 cell lysates. ICC/IF: PC-12 cells. IHC-P: Human breast 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid

  • Storage instructions

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

  • Dissociation constant (KD)

    KD = 6.03 x 10 -10 M
    Learn more about KD

  • Storage buffer

    pH: 7.20
    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

    EPR6301

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab124670 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
ICC/IF 1/500.
WB 1/10000 - 1/50000. Predicted molecular weight: 24 kDa.
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Target

Images

  • Western blot - Anti-HMGB2 antibody [EPR6301] (ab124670)
    Western blot - Anti-HMGB2 antibody [EPR6301] (ab124670)
    All lanes : Anti-HMGB2 antibody [EPR6301] (ab124670) at 1/2000 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 - ab124670 observed at 24 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab124670 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. ab124670 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 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 [EPR6301] (ab124670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGB2 antibody [EPR6301] (ab124670)

    ab124670, at a 1/250 dilution, staining HMGB2 in paraffin embedded Human breast tissue by Immunohistochemistry.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-HMGB2 antibody [EPR6301] (ab124670)
    Immunocytochemistry/ Immunofluorescence - Anti-HMGB2 antibody [EPR6301] (ab124670)

    Immunocytochemistry/Immunofluorescence analysis of PC-12 cells labelling HMGB2 with ab124670 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
    Control: PBS only.
    Nuclear counter stain: DAPI.

  • Western blot - Anti-HMGB2 antibody [EPR6301] (ab124670)
    Western blot - Anti-HMGB2 antibody [EPR6301] (ab124670)
    All lanes : Anti-HMGB2 antibody [EPR6301] (ab124670) at 1/10000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : HMGB2 knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : K562 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 24 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab124670 observed at 24 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab124670 was shown to specifically react with HMGB2 in wild-type HAP1 cells as signal was lost in HMGB2 knockout cells. Wild-type and HMGB2 knockout samples were subjected to SDS-PAGE. Ab124670 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 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.

  • Western blot - Anti-HMGB2 antibody [EPR6301] (ab124670)
    Western blot - Anti-HMGB2 antibody [EPR6301] (ab124670)
    All lanes : Anti-HMGB2 antibody [EPR6301] (ab124670) at 1/10000 dilution

    Lane 1 : K562 cell lysates
    Lane 2 : HeLa cell lysates
    Lane 3 : PC12 cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat-anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 24 kDa

  • OI-RD Scanning - Anti-HMGB2 antibody [EPR6301] (ab124670)
    OI-RD Scanning - Anti-HMGB2 antibody [EPR6301] (ab124670)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-HMGB2 antibody [EPR6301] (ab124670)
    Anti-HMGB2 antibody [EPR6301] (ab124670)

References (6)

Publishing research using ab124670? Please let us know so that we can cite the reference in this datasheet.

ab124670 has been referenced in 6 publications.

  • Pu J  et al. Long Noncoding RNA PART1 Promotes Hepatocellular Carcinoma Progression via Targeting miR-590-3p/HMGB2 Axis. Onco Targets Ther 13:9203-9211 (2020). PubMed: 32982307
  • Zhang W  et al. 6-Gingerol Attenuates Ischemia-Reperfusion-Induced Cell Apoptosis in Human AC16 Cardiomyocytes through HMGB2-JNK1/2-NF-?B Pathway. Evid Based Complement Alternat Med 2019:8798653 (2019). PubMed: 30886640
  • Zhang Q  et al. iTRAQ-based proteomic analysis of endotoxin tolerance induced by lipopolysaccharide. Mol Med Rep 20:584-592 (2019). PubMed: 31115557
  • Zhang P  et al. High-mobility group box 2 promoted proliferation of cervical cancer cells by activating AKT signaling pathway. J Cell Biochem 120:17345-17353 (2019). PubMed: 31209930
  • Davies JE  et al. Cross-reactivity of anti-HMGB1 antibodies for HMGB2. J Immunol Methods 456:72-76 (2018). PubMed: 29453955
  • He YH  et al. Association of Serum HMGB2 Levels With In-Stent Restenosis: HMGB2 Promotes Neointimal Hyperplasia in Mice With Femoral Artery Injury and Proliferation and Migration of VSMCs. Arterioscler Thromb Vasc Biol 37:717-729 (2017). PubMed: 28183701

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-HMGB2 antibody [EPR6301]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Lung)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
25 µg
Specification
Lung
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 7.5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Oct 16 2018

ChIP abreview for Anti-HMGB2 antibody [EPR6301]

 Good
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - nuclear (HUVEC)
Negative control
olfactory region 8 (OR8)
Specification
HUVEC
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 60 minute(s) and 0 second(s)
Specification of the cross-linking agent: 20 min EGS + 40 min with PFA w
Positive control
5’ prime UTR of ZNF335 and HIST2H2AC genes
Read More

Dr. Theodore Georgomanolis

Verified customer

Submitted May 25 2016

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