Product Information

Contents: Carrier-Free Recombinant Mouse HMGB1 (high mobility group box 1)
Catalog Number: 34-8401
Sizes: 100 ug, 500 ug
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.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
34-8401	Carrier-Free Recombinant Mouse HMGB1 (high mobility group box 1)	N/A	N/A	BA ELISA std

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Description

High-mobility group box-1 (HMGB1) 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 HMGB1 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). The cytokine activity of HMGB1 is contained in the B box, while the A box is associated with the helix-loop-helix domain of transcription factors. HMGB1 acts both as an inflammatory mediator that promotes monocyte migration and cytokine secretion, as well as a mediator of T cell-dendritic cell interaction. HMGB1 is secreted and acts to transduce cellular signals through its high affinity receptor, RAGE and possibly, TLR2 and TLR4.

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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