Contents: Carrier-Free Recombinant Mouse HMGB1 (high mobility group box 1)
Catalog Number: 34-8401
Formulation: Sterile liquid; 20 mM Tris-HCl, pH 8.0, 0.3 M NaCl, 1 mM DTT. 0.22 um filtered.
Storage Conditions: For greatest stability, keep concentration of primary stock at or above 10 µg/ml. For long term storage, aliquot into polypropylene vials (volumes of 20 µl or greater) and store at or below -80°C. Avoid repeated freeze/thaw cycles.
Handling Conditions: For best recovery, always quick-spin vial prior to opening. For dilution of current stock, always include carrier protein (1% BSA or 10% FBS) in the buffered saline diluent.
Source: E. coli expressed amino acids Met1-Glu215 of mouse HMGB-1 (accession # NM_010439).
Molecular Mass: The protein has a predicted molecular mass of 24,893. The DTT reduced protein migrates as a 30 kDa polypeptide on SDS-PAGE. The non-reduced protein migrates as a 30 kDa polypeptide.
Purity: Greater than 98%, as determined by SDS-PAGE
Endotoxin Level: Less than 0.01 ng/ug cytokine as determined by the LAL assay.
Bioactivity: Measured by its ability to induce TNF-a secretion by RAW264.7, a mouse macrophage cell line, in the presence of 10 µg/mL of polymyxin B. The ED50 for this effect is typically 3 - 12 µg/mL.
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Description
High-mobility group box-1 (HMGB-1) protein was originally described as a nuclear non-histone DNA binding chromosomal protein. However, recent studies indicate that damaged, necrotic cells liberate HMGB1 into the extracellular milieu where it functions as a proinflammatory cytokine. Mouse HMGB-1 is expressed as a 215 amino acid single chain polypeptide containing three domains: two tandem-linked positively charged DNA-binding domains (HMG box A, aa 9-79; and box B, aa 89-162), and a negatively charged 30 aa C-terminal acidic tail region. Residues 28 - 44 and 180 - 185 contain a nuclear localization signal (NLS). HMGB-1 is expressed at high levels in almost all cells. It 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. The cytokine activity of HMGB-1 is contained in the B box, while the A box is associated with the helix-loop-helix domain of transcription factors. HMBG-1 is released in response to cell death and as a secretion product. Once secreted, HMGB-1 transduces cellular signals through its high affinity receptor, RAGE and possibly, TLR2 and TLR4. Mouse HMGB1 is 99% aa identical to human HMGB.
Applications Reported
For research use only, not for diagnostic or therapeutic use. The recombinant mouse HMGB-1 has been reported useful for bioassay and ELISA.
Applications Tested
The recombinant mouse HMGB-1 has been tested by bioassay, yielding ED50 of 3 µg/mL.
References
Yang D, Chen Q, Yang H, Tracey KJ, Bustin M, Oppenheim JJ. High mobility group box-1 protein induces the migration and activation of human dendritic cells and acts as an alarmin. J Leukoc Biol. 2007, 81(1):59-66.
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