Human HMGB1/HMG-1 Antibody

(16 citations)
(2 Reviews)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human HMGB1/HMG-1 in direct ELISAs and Western blots.
  • Source
    Monoclonal Mouse IgG2B Clone # 115603
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human HMGB1/HMG-1
    Gly2-Glu215 (predicted)
    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
    0.05 µg/mL
    See below
  • Chromatin Immunoprecipitation (ChIP)
    5 µg/5 x 106 cells
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    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 HMGB1/HMG‑1 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, Jurkat human acute T cell leukemia cell line, and HEK293 human embryonic kidney cell line. PVDF membrane was probed with 0.05 µg/mL of Mouse Anti-Human HMGB1/HMG‑1 Monoclonal Antibody (Catalog # MAB1690) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for HMGB1/HMG‑1 at approximately 26 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Chromatin Immunoprecipitation (ChIP)
Detection of HMGB1/HMG‑1-regulated Genes by Chromatin Immunoprecipitation. HepG2 human hepatocellular carcinoma cell line was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. HMGB1/HMG‑1/DNA complexes were immunoprecipitated using 5 μg Mouse Anti-Human HMGB1/HMG‑1 PE-conjugated Monoclonal Antibody (Catalog # MAB1690) or control antibody (Catalog # MAB004) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Mouse IgG Secondary Antibody (Catalog # BAF007). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The AFP promoter was detected by standard PCR.
Intracellular Staining by Flow Cytometry
Detection of HMGB1 in HCT‑116 Human Cell Line by Flow Cytometry. HCT‑116 human colorectal carcinoma cell line was stained with Mouse Anti-Human HMGB1/HMG-1 Monoclonal Antibody (Catalog # MAB1690, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Immunocytochemistry
HMGB1/HMG‑1 in CCD‑18Co Human Cell Line. HMGB1/HMG‑1 was detected in immersion fixed CCD‑18Co human colonic fibroblast cell line using Mouse Anti-Human HMGB1/HMG‑1 Monoclonal Antibody (Catalog # MAB1690) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
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.

16 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Anti-high mobility group box-1 (HMGB1) antibody attenuates delayed cerebral vasospasm and brain injury after subarachnoid hemorrhage in rats
    Sci Rep, 2016;6(0):37755.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  2. Expression of RAGE and HMGB1 in thymic epithelial tumors, thymic hyperplasia and regular thymic morphology.
    Authors: Moser B, Janik S, Schiefer A, Mullauer L, Bekos C, Scharrer A, Mildner M, Renyi-Vamos F, Klepetko W, Ankersmit H
    PLoS ONE, 2014;9(4):e94118.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  3. A dual-role of Gu-4 in suppressing HMGB1 secretion and blocking HMGB1 pro-inflammatory activity during inflammation.
    Authors: Zhou H, Ji X, Wu Y, Xuan J, Qi Z, Shen L, Lan L, Li Q, Yin Z, Li Z, Zhao Z
    PLoS ONE, 2014;9(3):e89634.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
  4. Local and systemic RAGE axis changes in pulmonary hypertension: CTEPH and iPAH.
    Authors: Moser B, Megerle A, Bekos C, Janik S, Szerafin T, Birner P, Schiefer A, Mildner M, Lang I, Skoro-Sajer N, Sadushi-Kolici R, Taghavi S, Klepetko W, Ankersmit H
    PLoS ONE, 2014;9(9):e106440.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  5. IL-1alpha released from damaged epithelial cells is sufficient and essential to trigger inflammatory responses in human lung fibroblasts.
    Authors: Suwara M, Green N, Borthwick L, Mann J, Mayer-Barber K, Barron L, Corris P, Farrow S, Wynn T, Fisher A, Mann D
    Mucosal Immunol, 2014;7(3):684-93.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
  6. Measles virus causes immunogenic cell death in human melanoma.
    Authors: Donnelly, O G, Errington-Mais, F, Steele, L, Hadac, E, Jennings, V, Scott, K, Peach, H, Phillips, R M, Bond, J, Pandha, H, Harrington, K, Vile, R, Russell, S, Selby, P, Melcher, A A
    Gene Ther, 2013;20(1):7-15.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: WB
  7. Sputum biomarkers and the prediction of clinical outcomes in patients with cystic fibrosis.
    Authors: Liou T, Adler F, Keogh R, Li Y, Jensen J, Walsh W, Packer K, Clark T, Carveth H, Chen J, Rogers S, Lane C, Moore J, Sturrock A, Paine R, Cox D, Hoidal J
    PLoS ONE, 2012;7(8):e42748.
    Species: Human
    Sample Type: Sputum
    Application: ELISA Development
  8. Human Papillomavirus Type 8 Interferes with a Novel C/EBPβ-Mediated Mechanism of Keratinocyte CCL20 Chemokine Expression and Langerhans Cell Migration.
    Authors: Sperling T, Oldak M, Walch-Ruckheim B, Wickenhauser C, Doorbar J, Pfister H, Malejczyk M, Majewski S, Keates AC, Smola S
    PLoS Pathog., 2012;8(7):e1002833.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  9. Fecal HMGB1 is a novel marker of intestinal mucosal inflammation in pediatric inflammatory bowel disease.
    Authors: Vitali R, Stronati L, Negroni A
    Am. J. Gastroenterol., 2011;106(11):2029-40.
    Species: Human
    Sample Type: Feces
    Application: WB
  10. Ethyl pyruvate decreases HMGB1 release and ameliorates murine colitis.
    Authors: Dave SH, Tilstra JS, Matsuoka K, Li F, DeMarco RA, Beer-Stolz D, Sepulveda AR, Fink MP, Lotze MT, Plevy SE
    J. Leukoc. Biol., 2009;86(3):633-43.
    Species: Mouse
    Sample Type: Feces
    Application: ELISA Development
  11. Functional dissection of an IFN-alpha/beta receptor 1 promoter variant that confers higher risk to chronic hepatitis B virus infection.
    Authors: Zhou J, Huang JD, Poon VK, Chen DQ, Chan CC, Ng F, Guan XY, Watt RM, Lu L, Yuen KY, Zheng BJ
    J. Hepatol., 2009;51(2):322-32.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  12. Potential role of high-mobility group box 1 in cystic fibrosis airway disease.
    Authors: Rowe SM, Jackson PL, Liu G, Hardison M, Livraghi A, Solomon GM, McQuaid DB, Noerager BD, Gaggar A, Clancy JP, O'Neal W, Sorscher EJ, Abraham E, Blalock JE
    Am. J. Respir. Crit. Care Med., 2008;178(8):822-31.
    Species: Human
    Sample Type: Saliva
    Application: WB
  13. E-selectin regulates gene expression in metastatic colorectal carcinoma cells and enhances HMGB1 release.
    Authors: Aychek T, Miller K, Sagi-Assif O, Levy-Nissenbaum O, Israeli-Amit M, Pasmanik-Chor M, Jacob-Hirsch J, Amariglio N, Rechavi G, Witz IP
    Int. J. Cancer, 2008;123(8):1741-50.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Immunodepletion
  14. TLR4 enhances TGF-beta signaling and hepatic fibrosis.
    Authors: Seki E, De Minicis S, Osterreicher CH, Kluwe J, Osawa Y, Brenner DA, Schwabe RF
    Nat. Med., 2007;13(11):1324-32.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  15. Pivotal advance: inhibition of HMGB1 nuclear translocation as a mechanism for the anti-rheumatic effects of gold sodium thiomalate.
    Authors: Zetterstrom CK, Jiang W, Wahamaa H, Ostberg T, Aveberger AC, Schierbeck H, Lotze MT, Andersson U, Pisetsky DS, Erlandsson Harris H
    J. Leukoc. Biol., 2007;83(1):31-8.
    Species: Mouse
    Sample Type: Cell Culture Supernates
    Application: WB
  16. IL-1alpha and IL-1beta are endogenous mediators linking cell injury to the adaptive alloimmune response.
    Authors: Rao DA, Tracey KJ, Pober JS
    J. Immunol., 2007;179(10):6536-46.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
Expand to show all 16 Citations
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Average Rating: 4 (Based on 2 reviews)

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We have 2 reviews tested in 2 species: Human, Mouse.
We have 2 reviews tested in 2 applications: Immunocytochemistry/Immunofluorescence, Western Blot.

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Images Ratings Applications Species Reviewed By Date Details
Immunocytochemistry/Immunofluorescence Human HMGB1/HMG-1 Antibody MAB1690
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Excellent
 ICC/IF Mouse Anonymous 10/13/2016
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Image Details
Immunocytochemistry/Immunofluorescence Human HMGB1/HMG-1 Antibody MAB1690
Immunocytochemistry/Immunofluorescence: Human HMGB1/HMG-1 Antibody [MAB1690]

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample Testedfibrotic lung
SpeciesMouse
  Average
 WB Human Anonymous 01/23/2016
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Summary

ApplicationWestern Blot
Sample TestedCell lysate
SpeciesHuman

Other Experimental Details

Other Experimental DetailsI should say this is a good product. I purchased the small size, detected HMGB1 expression after TNF-a challenged. It is helpful.

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