Human HMGB1/HMG-1 Alexa Fluor® 700-conjugated Antibody

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FAB1690N-100UG
R&D Systems Antibodies
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Human HMGB1/HMG-1 Alexa Fluor® 700-conjugated Antibody Summary

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
Supplied 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.
Label
Alexa Fluor 700 (Excitation= 675-700 nm, Emission= 723 nm)

Applications

Recommended Concentration
Sample
Intracellular Staining by Flow Cytometry
0.25-1 µg/106 cells
HCT‑116 human colorectal carcinoma cell line fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

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.

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

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

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Product Specific Notices


This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

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