ARG70220

Human HMGB1 recombinant protein (Active) (His-tagged, C-ter)

Human HMGB1 recombinant protein (Active) (His-tagged, C-ter) for Functional study,SDS-PAGE and Human,Mouse

Overview

Product Description E. coli expressed, His-tagged (C-ter) Active Human HMGB1 recombinant protein.
Tested Reactivity Hu, Ms
Tested Application FuncSt, SDS-PAGE
Target Name HMGB1
Species Human
A.A. Sequence MGKGD PKKPR GKMSS YAFFV QTCRE EHKKK HPDAS VNFSE FSKKC SERWK TMSAK EKGKF EDMAK ADKAR YEREM KTYIP PKGET KKKFK DPNAP KRPPS AFFLF CSEYR PKIKG EHPGL SIGDV AKKLG EMWNN TAADD KQPYE KKAAK LKEKY EKDIA AYRAK GKPDA AKKGV VKAEK SKKKK EEEED EEDEE DEEEE EDEED EDEEE DDDDE with polyhistidine tag at the C-terminus.
Expression System E. coli
Activity Active
Activity Note Measure by its ability to induce TNF alpha in Raw264.7 cells.
The ED50 for this effect is < 10 μg/ml. This protein also induces TNF alpha releasing from human A549 cultured cells based on customer feedback.
Alternate Names HMG-1; High mobility group protein B1; High mobility group protein 1; HMG1; SBP-1; HMG3

Properties

Form Powder
Purification Ni-NTA chromatography.
Purification Note Endotoxin level is less than 0.1 EU/µg of the protein, as determined by the LAL test.
Purity > 98% (by SDS-PAGE)
Buffer PBS (pH 8.0)
Reconstitution It is recommended to reconstitute the lyophilized protein in sterile water to a concentration not less than 100 µg/ml and incubate the stock solution for at least 20 min at room temperature to make sure the protein is dissolved completely.
Storage Instruction Lyophilized protein should be stored at -20°C. After reconstitution, aliquot and store at -20°C or -80°C for up to one month. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening.
Note For laboratory research only, not for drug, diagnostic or other use.

Bioinformation

Database Links

GeneID: 15289 Mouse HMGB1

GeneID: 3146 Human HMGB1

Swiss-port # P09429 Human High mobility group protein B1

Swiss-port # P63158 Mouse High mobility group protein B1

Gene Symbol HMGB1
Gene Full Name high mobility group box 1
Background HMGB1 is a protein that belongs to the High Mobility Group-box superfamily. The encoded non-histone, nuclear DNA-binding protein regulates transcription, and is involved in organization of DNA. This protein plays a role in several cellular processes, including inflammation, cell differentiation and tumor cell migration. Multiple pseudogenes of this gene have been identified. Alternative splicing results in multiple transcript variants that encode the same protein. [provided by RefSeq, Sep 2015]
Function HMGB1 is a DNA binding protein. It associates with chromatin and has the ability to bend DNA. Binds preferentially single-stranded DNA. Involved in V(D)J recombination by acting as a cofactor of the RAG complex. Acts by stimulating cleavage and RAG protein binding at the 23 bp spacer of conserved recombination signal sequences (RSS). [UniProt]
Cellular Localization Nucleus. Chromosome. Cytoplasm. Secreted. Cell membrane; Peripheral membrane protein; Extracellular side. Endosome. Endoplasmic reticulum-Golgi intermediate compartment. Note=In basal state predominantly nuclear. Shuttles between the cytoplasm and the nucleus. Translocates from the nucleus to the cytoplasm upon autophagy stimulation. Release from macrophages in the extracellular milieu requires the activation of NLRC4 or NLRP3 inflammasomes. [UniProt]
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Calculated MW 25 kDa
PTM Phosphorylated at serine residues. Phosphorylation in both NLS regions is required for cytoplasmic translocation followed by secretion (PubMed:17114460).

Acetylated on multiple sites upon stimulation with LPS (PubMed:22801494). Acetylation on lysine residues in the nuclear localization signals (NLS 1 and NLS 2) leads to cytoplasmic localization and subsequent secretion (By similarity). Acetylation on Lys-3 results in preferential binding to DNA ends and impairs DNA bending activity (By similarity).

Reduction/oxidation of cysteine residues Cys-23, Cys-45 and Cys-106 and a possible intramolecular disulfide bond involving Cys-23 and Cys-45 give rise to different redox forms with specific functional activities in various cellular compartments: 1- fully reduced HMGB1 (HMGB1C23hC45hC106h), 2- disulfide HMGB1 (HMGB1C23-C45C106h) and 3- sulfonyl HMGB1 (HMGB1C23soC45soC106so).

Poly-ADP-ribosylated by PARP1 when secreted following stimulation with LPS (By similarity).

In vitro cleavage by CASP1 is liberating a HMG box 1-containing peptide which may mediate immunogenic activity; the peptide antagonizes apoptosis-induced immune tolerance (PubMed:24474694). Can be proteolytically cleaved by a thrombin:thrombomodulin complex; reduces binding to heparin and proinflammatory activities (By similarity). [UniProt]