Product Specifications for Recombinant Human HMGB1 Protein, CF
Mouse myeloma cell line, NS0-derived human HMGB1/HMG-1 protein Met1-Glu215
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Met1 & Gly2
Predicted Molecular Mass
28-34 kDa, reducing conditions
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
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant
protein to be stored at a more dilute concentration.
The carrier free version does not contain BSA.
What formulation is right for me?
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or
as an ELISA standard.
In contrast, the carrier free protein is recommended for applications, in which the presence of BSA
Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitute at 200 μg/mL in PBS.
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.
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).
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High Mobility Group Protein 1
Entrez Gene IDs
Amphoterin, HMG-1, HMG1, HMG1DKFZp686A04236, HMG3, HMGB1, High mobility group protein 1, SBP-1, Sulfoglucuronyl carbohydrate binding protein, high mobility group box 1, high mobility group protein B1, high-mobility group (nonhistone chromosomal) protein 1, high-mobility group box 1
Which Brands are Currently Available on bio‑techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments are available to quote. ACD branded products will be available on bio‑techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
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