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Recombinant Human HMGB1 Protein, CF Best Seller

Catalog # 1690-HMB | R&D Systems, Inc. a Bio-Techne Brand
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1690-HMB-050

Key Product Details

Source

NS0

Accession #

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human HMGB1/HMG-1 protein
Met1-Glu215

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met1 & Gly2

Predicted Molecular Mass

25 kDa

SDS-PAGE

28-34 kDa, reducing conditions

Activity

Measured by its ability to induce TNF-alpha secretion by RAW 264.7 mouse monocyte/macrophage cells.
The ED50 for this effect is 2.5-12.5 μg/mL.

Formulation, Preparation and Storage

1690-HMB
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 200 μg/mL in PBS.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: HMGB1/HMG-1

High-mobility Group Box 1 protein (HMGB1), also known as HMG1 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 25 kDa single chain protein containing two globular positively charged DNA-binding domains (HMG boxes A and B) and a negatively charged C-terminal region that contains only Asp and Glu residues (4, 5). Posttranslational modification of HMGB1, including acetylation, phosphorylation, and methylation, affects HMGB1 localization, receptor interactions, and bioactivity (3). An intramolecular disulfide bond between Cys23 and Cys45 as well as the presence of the unpaired Cys106 thiol are critical for HMGB1-induced pro-inflammatory TNF-alpha secretion (2, 6). Alternatively, fully reduced HMGB1 acts as a potent chemoattractant for neutrophils and monocytes (7). HMGB1 can be localized to the nucleus or cytoplasm and can also be secreted despite its lack of a signal peptide (2). HMGB1 binds DNA in a non-sequence specific manner and may act as a structural cofactor during gene transcription (8). Acetylation of HMGB1 results in its cytoplasmic localization and eventual secretion (9). HMGB1 can be secreted by multiple cell types, and it is also released upon cell necrosis, apoptosis, and pyroptosis (2, 3). HMGB1 is widely recognized as a multifunctional alarmin that stimulates inflammation upon sterile or infectious insult. Receptors for HMGB1 include TLR2, TLR4, TLR9, Syndecan-3, Siglec-10, Integrin alpha M beta 2, CXCR4, TIM-3, and RAGE (1, 2). It is implicated in the pathogenesis of a broad range of diseases including atherosclerosis, sepsis, cancer, neurodegenerative diseases, and autoimmunity (3, 10-13).

References

  1. Yanai, H. et al. (2012) Trends Immunol. 33:633.
  2. Yang, H. et al. (2013) J. Leukoc. Biol. 93:865.
  3. Harris, H.E. et al. (2012) Nat. Rev. Rheumatol. 8:195.
  4. Degryse, B. and M. de Virgilio (2003) FEBS Lett. 553:11.
  5. Wen, L. et al. (1989) Nucleic Acids Res. 17:1197.
  6. Yang, H. et al. (2021) Mol. Med. 27:58.
  7. Venereau, E. et al. (2012) J. Exp. Med. 209:1519.
  8. Bianchi, M.E. and A. Agresti (2005) Curr. Opin. Genet. Dev. 15:496.
  9. Bonaldi, T. et al. (2003) EMBO J. 22:5551.
  10. Diener, K.R. et al. (2013) Immunol. Cell Biol. 91:443.
  11. Fang, P. et al. (2012) Mol. Neurobiol. 45:499.
  12. de Souza, A.W. et al. (2012) Autoimmun. Rev. 11:909.
  13. Bae, J.S. (2012) Arch. Pharm. Res. 35:1511.

Long Name

High Mobility Group Protein 1

Alternate Names

HMG-1, HMG1

Entrez Gene IDs

3146 (Human)

Gene Symbol

HMGB1

UniProt

Product Documents for Recombinant Human HMGB1 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human HMGB1 Protein, CF

For research use only

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FAQs for Recombinant Human HMGB1 Protein, CF

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  • Q: What is the oxidation state of Recombinant Human HMGB1 Protein, CF (Catalog # 1690-HMB)?

    A: The presence of both a disulfide bond between Cys23 and Cys45, as well as an unpaired Cys106 thiol, has been reported in the literature to be critical for HMBG1-induced TNF-alpha secretion. The activity of 1690-HMB is measured by its ability to induce TNF-alpha secretion by RAW 264.7 mouse monocyte/macrophage cells. Each new bulk lot of 1690-HMB must exhibit acceptable activity in the TNF-alpha secretion assay to pass our activity specifications. The oxidiation state of 1690-HMB is not a product specification and has not been fully characterized in-house.

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