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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-HMGB2 antibody [EPR6301] (ab124670)

Reviews (2) Submit a question References (10)
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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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.

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

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR6301

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

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)
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.
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 (10)

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

ab124670 has been referenced in 10 publications.

  • Lüdtke TH  et al. Combined genomic and proteomic approaches reveal DNA binding sites and interaction partners of TBX2 in the developing lung. Respir Res 22:85 (2021). PubMed: 33731112
  • Han X  et al. Identification of differentially expressed proteins and clinicopathological significance of HMGB2 in cervical cancer. Clin Proteomics 18:2 (2021). PubMed: 33407071
  • Li Y  et al. microRNA-130a-5p suppresses myocardial ischemia reperfusion injury by downregulating the HMGB2/NF-?B axis. BMC Cardiovasc Disord 21:121 (2021). PubMed: 33658008
  • 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
  • Olins AL  et al. Hyperosmotic stress: in situ chromatin phase separation. Nucleus 11:1-18 (2020). PubMed: 31924112
  • 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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