Human/Mouse/Rat HMGB1/HMG-1 Antibody

(1 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat HMGB1/HMG-1 in Western blot.
  • Source
    Monoclonal Rat IgG2B Clone # 951420
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Human HMGB1/HMG-1 synthetic peptide
    Accession # P09429
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Immunohistochemistry
    5-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human, Mouse, and Rat HMGB1/HMG‑1 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, Hepa 1‑6 mouse hepatoma cell line, H4‑II‑E‑C3 rat hepatoma cell line, HEK293 human embryonic kidney cell line, and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 0.1 µg/mL of Rat Anti-Human HMGB1/HMG‑1 Monoclonal Antibody (Catalog # MAB16901) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for HMGB1/HMG‑1 at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
HMGB1/HMG‑1 in Human Prostate Cancer Tissue. HMGB1/HMG‑1 was detected in immersion fixed paraffin-embedded sections of human prostate cancer tissue using Rat Anti-Human/Mouse/Rat HMGB1/HMG‑1 Monoclonal Antibody (Catalog # MAB16901) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS017) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei and cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: HMGB1/HMG-1

Human High-mobility group box 1 protein (HMGB1), previously known as HMG-1 or amphoterin, is a member of the high mobility group box family of non-histone chromosomal proteins (1‑3). Human HMGB1 is expressed as a 30 kDa, 215 amino acid (aa) single chain polypeptide containing three domains: two N-terminal globular, 70 aa positively charged DNA-binding domains (HMG boxes A and B), and a negatively charged 30 aa C-terminal region that contains only Asp and Glu (4, 5). Residues 27‑43 and 178‑184 contain a NLS. Posttranslational modifications of the molecule have been reported, with acetylation occurring on as many as 17 lysine residues (6). HMGB1 is expressed at high levels in almost all cells (2, 4). It was originally discovered as a nuclear protein that could bend DNA. Such bending stabilizes nucleosome formation and regulates the expression of select genes upon recruitment by DNA binding proteins (1, 7, 8). It is now known that HMGB1 can also act extracellularly, both as an inflammatory mediator that promotes monocyte migration and cytokine secretion, and as a mediator of T cell-dendritic cell interaction (1, 4, 7, 9, 10). The cytokine activity of HBMG1 is restricted to the HMG B box, (3) while the A box is associated with the helix-loop-helix domain of transcription factors (11). HMBG1 is released in response to cell death and as a secretion product. Although HMBG-1 does not possess a classic signal sequence, it appears to be secreted as an acetylated form via secretory endolysosome exocytosis (6, 12). Once secreted, HMGB1 transduces cellular signals through its high affinity receptor, RAGE and, possibly, TLR2 and TLR4 (1, 3, 4). Human HMGB1 is 100% aa identical to canine HMGB1 and 99% aa identical to mouse, rat, bovine and porcine HMGB1, respectively.

  • References:
    1. Lotze, M.T. and K.J. Tracey (2005) Nat. Rev. Immunol. 5:331.
    2. Yang, H. et al. (2005) J. Leukoc. Biol. 78:1.
    3. Dumitriu, I.E. et al. (2005) Trends Immunol. 26:381.
    4. Degryse, B. and M. de Virgilio (2003) FEBS Lett. 553:11.
    5. Wen, L. et al. (1989) Nucleic Acids Res. 17:1197.
    6. Bonaldi, T. et al. (2003) EMBO J. 22:5551.
    7. Muller, S. et al. (2001) EMBO J. 20:4337.
    8. Bustin, M. (1999) Mol. Cell. Biol. 19:5237.
    9. Wang, H. et al. (1999) Science. 285:248.
    10. Dimitriu, I.E. et al. (2005) J. Immunol. 174:7506.
    11. Najima, Y. et al. (2005) J. Biol. Chem. 280:27523.
    12. Gardella, S. et al. (2002) EMBO Rep. 3:995.
  • Long Name:
    High Mobility Group Protein 1
  • Entrez Gene IDs:
    3146 (Human)
  • Alternate Names:
    Amphoterin; high mobility group box 1; High mobility group protein 1; high mobility group protein B1; high-mobility group (nonhistone chromosomal) protein 1; high-mobility group box 1; HMG1; HMG-1; HMG1DKFZp686A04236; HMG3; HMGB1; SBP-1; Sulfoglucuronyl carbohydrate binding protein
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. Anti-high mobility group box-1 (HMGB1) antibody inhibits hemorrhage-induced brain injury and improved neurological deficits in rats
    Authors: D Wang, K Liu, H Wake, K Teshigawar, S Mori, M Nishibori
    Sci Rep, 2017;7(0):46243.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Rat HRP-DAB Cell & Tissue Staining Kit

CTS017 5
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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

CTS013 4
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Mounting Medium

CTS011 4
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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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Isotype Controls
Description Application Cat# Citations Images  

Rat IgG2B Isotype Control

Ctrl MAB0061 27  
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Normal Rat IgG Control

Ctrl 6-001-A 9
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Normal Rat IgG Control (Azide Free)

Ctrl 6-001-F 1
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Rat IgG2B Isotype Control

Ctrl MAB0061R
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Secondary Antibodies
Description Application Cat# Citations Images  

Rat IgG HRP-conjugated Antibody

WB, Simple Western HAF005 7  
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Goat Anti-Rat F(ab)2 IgG (H+L)
APC-conjugated Antibody

Flow F0113 2  
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Goat Anti-Rat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL013 2
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Rat F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0105B 2  
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Goat Anti-Rat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL015 1
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Goat Anti-Rat IgG Biotinylated Antibody

WB BAF005 2
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Rat IgG VisUCyte HRP Polymer Antibody

IHC VC005  
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Goat Anti-Rat IgG Antibody

WB AF005 1
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Rat F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0104B  
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Goat Anti-Rat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL014
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Goat Anti-Rat F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0115 1
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