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Epigenetics and Nuclear Signaling Histones HMGs
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Recombinant human HMGB1 protein (ab167718)

  • Datasheet
  • SDS
Reviews (2)Q&A (1)References (2)

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SDS-PAGE - Recombinant Human HMGB1 protein (ab167718)
  • Functional Studies - Recombinant Human HMGB1 protein (ab167718)

Key features and details

  • Expression system: HEK 293 cells
  • Purity: > 95% SDS-PAGE
  • Endotoxin level: < 1.000 Eu/µg
  • Active: Yes
  • Tags: His tag C-Terminus
  • Suitable for: SDS-PAGE

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Description

  • Product name

    Recombinant human HMGB1 protein
    See all HMGB1 proteins and peptides
  • Biological activity

    Human HMGB1 Protein, His Tag stimulates the production of TNF-α in Raw-246.7 cells. The EC50 for this effect is 0.7855-0.8342 μg/ml (Routinely tested).

  • Purity

    > 95 % SDS-PAGE.

  • Endotoxin level

    < 1.000 Eu/µg
  • Expression system

    HEK 293 cells
  • Accession

    P09429
  • Protein length

    Protein fragment
  • Animal free

    No
  • Nature

    Recombinant
    • Species

      Human
    • Sequence

      GKGDPKKPRGKMSSYAFFVQTCREEHKKKHPDASVNFSEFSKKCSERWKT MSAKEKGKFEDMAKADKARYEREMKTYIPPKGETKKKFKDPNAPKRPPSA FFLFCSEYRPKIKGEHPGLSIGDVAKKLGEMWNNTAADDKQPYEKKAAKL KEKYEKDIAAYRAKGKPDAAKKGVVKAEKSKKKKEEEEDEEDEEDEEEEE DEEDEDEEEDDDDE
    • Predicted molecular weight

      26 kDa including tags
    • Amino acids

      2 to 215
    • Tags

      His tag C-Terminus

Associated products

  • Related Products

    • Anti-6X His tag® antibody [HIS.H8] (ab18184)
    • Anti-HMGB1 antibody (ab18256)
    • Anti-6X His tag® antibody [4D11] (ab5000)
    • Anti-HMGB1 antibody (ab77302)
    • Anti-HMGB1 antibody [EPR3507] (ab79823)
    • Anti-6X His tag® antibody (ab9108)
    • Anti-HMGB1 antibody [EPR3506] (ab92310)

Specifications

Our Abpromise guarantee covers the use of ab167718 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Applications

    SDS-PAGE

  • Form

    Lyophilized
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

    pH: 7.40
    Constituent: 99% PBS

    Normally Mannitol or Trehalose are added as protectants before lyophilization.

    This product is an active protein and may elicit a biological response in vivo, handle with caution.

  • Reconstitution
    It is recommended to reconstitute the lyophilized protein in 100µl sterile deionized water to a final concentration of 1mg/ml. Solubilize for 30 to 60 minutes at room temperature with occasional gentle mixing. Carrier protein (0.1% HSA or BSA) is strongly recommended for further dilution and long term storage.

General Info

  • Alternative names

    • Amphoterin
    • Chromosomal protein, nonhistone, HMG1
    • DKFZp686A04236
    • High mobility group 1
    • High mobility group box 1
    • High mobility group protein 1
    • High mobility group protein B1
    • high-mobility group (nonhistone chromosomal) protein 1
    • HMG-1
    • HMG1
    • HMG3
    • HMGB 1
    • HMGB1
    • HMGB1_HUMAN
    • NONHISTONE CHROMOSOMAL PROTEIN HMG1
    • SBP 1
    • Sulfoglucuronyl carbohydrate binding protein
    see all
  • Function

    Multifunctional redox sensitive protein with various roles in different cellular compartments. In the nucleus is one of the major chromatin-associated non-histone proteins and acts as a DNA chaperone involved in replication, transcription, chromatin remodeling, V(D)J recombination, DNA repair and genome stability. Proposed to be an universal biosensor for nucleic acids. Promotes host inflammatory response to sterile and infectious signals and is involved in the coordination and integration of innate and adaptive immune responses. In the cytoplasm functions as sensor and/or chaperone for immunogenic nucleic acids implicating the activation of TLR9-mediated immune responses, and mediates autophagy. Acts as danger associated molecular pattern (DAMP) molecule that amplifies immune responses during tissue injury. Released to the extracellular environment can bind DNA, nucleosomes, IL-1 beta, CXCL12, AGER isoform 2/sRAGE, lipopolysaccharide (LPS) and lipoteichoic acid (LTA), and activates cells through engagement of multiple surface receptors. In the extracellular compartment fully reduced HMGB1 (released by necrosis) acts as a chemokine, disulfide HMGB1 (actively secreted) as a cytokine, and sulfonyl HMGB1 (released from apoptotic cells) promotes immunological tolerance (PubMed:23519706, PubMed:23446148, PubMed:23994764, PubMed:25048472). Has proangiogdenic activity (By similarity). May be involved in platelet activation (By similarity). Binds to phosphatidylserine and phosphatidylethanolamide (By similarity). Bound to RAGE mediates signaling for neuronal outgrowth (By similarity). May play a role in accumulation of expanded polyglutamine (polyQ) proteins such as huntingtin (HTT) or TBP (PubMed:23303669, PubMed:25549101).
    Nuclear functions are attributed to fully reduced HGMB1. Associates with chromatin and binds DNA with a preference to non-canonical DNA structures such as single-stranded DNA, DNA-containing cruciforms or bent structures, supercoiled DNA and ZDNA. Can bent DNA and enhance DNA flexibility by looping thus providing a mechanism to promote activities on various gene promoters by enhancing transcription factor binding and/or bringing distant regulatory sequences into close proximity (PubMed:20123072). May have an enhancing role in nucleotide excision repair (NER) (By similarity). However, effects in NER using in vitro systems have been reported conflictingly (PubMed:19446504, PubMed:19360789). May be involved in mismatch repair (MMR) and base excision repair (BER) pathways (PubMed:15014079, PubMed:16143102, PubMed:17803946). May be involved in double strand break repair such as non-homologous end joining (NHEJ) (By similarity). 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) (By similarity). In vitro can displace histone H1 from highly bent DNA (By similarity). Can restructure the canonical nucleosome leading to relaxation of structural constraints for transcription factor-binding (By similarity). Enhances binding of sterol regulatory element-binding proteins (SREBPs) such as SREBF1 to their cognate DNA sequences and increases their transcriptional activities (By similarity). Facilitates binding of TP53 to DNA (PubMed:23063560). Proposed to be involved in mitochondrial quality control and autophagy in a transcription-dependent fashion implicating HSPB1; however, this function has been questioned (By similarity). Can modulate the activity of the telomerase complex and may be involved in telomere maintenance.
    In the cytoplasm proposed to dissociate the BECN1:BCL2 complex via competitive interaction with BECN1 leading to autophagy activation (PubMed:20819940). Involved in oxidative stress-mediated autophagy (PubMed:21395369). Can protect BECN1 and ATG5 from calpain-mediated cleavage and thus proposed to control their proautophagic and proapoptotic functions and to regulate the extent and severity of inflammation-associated cellular injury (By similarity). In myeloid cells has a protective role against endotoxemia and bacterial infection by promoting autophagy (By similarity). Involved in endosomal translocation and activation of TLR9 in response to CpG-DNA in macrophages.
    In the extracellular compartment (following either active secretion or passive release) involved in regulation of the inflammatory response. Fully reduced HGMB1 (which subsequently gets oxidized after release) in association with CXCL12 mediates the recruitment of inflammatory cells during the initial phase of tissue injury; the CXCL12:HMGB1 complex triggers CXCR4 homodimerization (PubMed:22370717). Induces the migration of monocyte-derived immature dendritic cells and seems to regulate adhesive and migratory functions of neutrophils implicating AGER/RAGE and ITGAM (By similarity). Can bind to various types of DNA and RNA including microbial unmethylated CpG-DNA to enhance the innate immune response to nucleic acids. Proposed to act in promiscuous DNA/RNA sensing which cooperates with subsequent discriminative sensing by specific pattern recognition receptors (By similarity). Promotes extracellular DNA-induced AIM2 inflammasome activation implicating AGER/RAGE (PubMed:24971542). Disulfide HMGB1 binds to transmembrane receptors, such as AGER/RAGE, TLR2, TLR4 and probably TREM1, thus activating their signal transduction pathways. Mediates the release of cytokines/chemokines such as TNF, IL-1, IL-6, IL-8, CCL2, CCL3, CCL4 and CXCL10 (PubMed:12765338, PubMed:18354232, PubMed:19264983, PubMed:20547845, PubMed:24474694). Promotes secretion of interferon-gamma by macrophage-stimulated natural killer (NK) cells in concert with other cytokines like IL-2 or IL-12 (PubMed:15607795). TLR4 is proposed to be the primary receptor promoting macrophage activation and signaling through TLR4 seems to implicate LY96/MD-2 (PubMed:20547845). In bacterial LPS- or LTA-mediated inflammatory responses binds to the endotoxins and transfers them to CD14 for signaling to the respective TLR4:LY96 and TLR2 complexes (PubMed:18354232, PubMed:21660935, PubMed:25660311). Contributes to tumor proliferation by association with ACER/RAGE (By similarity). Can bind to IL1-beta and signals through the IL1R1:IL1RAP receptor complex (PubMed:18250463). Binding to class A CpG activates cytokine production in plasmacytoid dendritic cells implicating TLR9, MYD88 and AGER/RAGE and can activate autoreactive B cells. Via HMGB1-containing chromatin immune complexes may also promote B cell responses to endogenous TLR9 ligands through a B-cell receptor (BCR)-dependent and ACER/RAGE-independent mechanism (By similarity). Inhibits phagocytosis of apoptotic cells by macrophages; the function is dependent on poly-ADP-ribosylation and involves binding to phosphatidylserine on the cell surface of apoptotic cells (By similarity). In adaptive immunity may be involved in enhancing immunity through activation of effector T cells and suppression of regulatory T (TReg) cells (PubMed:15944249, PubMed:22473704). In contrast, without implicating effector or regulatory T-cells, required for tumor infiltration and activation of T-cells expressing the lymphotoxin LTA:LTB heterotrimer thus promoting tumor malignant progression (By similarity). Also reported to limit proliferation of T-cells (By similarity). Released HMGB1:nucleosome complexes formed during apoptosis can signal through TLR2 to induce cytokine production (PubMed:19064698). Involved in induction of immunological tolerance by apoptotic cells; its pro-inflammatory activities when released by apoptotic cells are neutralized by reactive oxygen species (ROS)-dependent oxidation specifically on Cys-106 (PubMed:18631454). During macrophage activation by activated lymphocyte-derived self apoptotic DNA (ALD-DNA) promotes recruitment of ALD-DNA to endosomes.
  • Tissue specificity

    Ubiquituous. Expressed in platelets (PubMed:11154118).
  • Sequence similarities

    Belongs to the HMGB family.
    Contains 2 HMG box DNA-binding domains.
  • Domain

    HMG box 2 mediates proinflammatory cytokine-stimulating activity and binding to TLR4 (PubMed:12765338, PubMed:20547845). However, not involved in mediating immunogenic activity in the context of apoptosis-induced immune tolerance (PubMed:24474694).
    The acidic C-terminal domain forms a flexible structure which can reversibly interact intramolecularily with the HMG boxes and modulate binding to DNA and other proteins (PubMed:23063560).
  • Post-translational
    modifications

    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.
    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.
    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.
  • Cellular localization

    Nucleus. Chromosome. Cytoplasm. Secreted. Cell membrane. Endosome. Endoplasmic reticulum-Golgi intermediate compartment. In basal state predominantly nuclear. Shuttles between the cytoplasm and the nucleus (PubMed:12231511, PubMed:17114460). Translocates from the nucleus to the cytoplasm upon autophagy stimulation (PubMed:20819940). Release from macrophages in the extracellular milieu requires the activation of NLRC4 or NLRP3 inflammasomes (By similarity). Passively released to the extracellular milieu from necrotic cells by diffusion, involving the fully reduced HGMB1 which subsequently gets oxidized (PubMed:19811284). Also released from apoptic cells (PubMed:16855214, PubMed:18631454). Active secretion from a variety of immune and non-immune cells such as macrophages, monocytes, neutrophils, dendritic cells and natural killer cells in response to various stimuli such as LPS and cytokines involves a nonconventional secretory process via secretory lysosomes (PubMed:12231511, PubMed:14532127, PubMed:15944249). Secreted by plasma cells in response to LPS (By similarity). Found on the surface of activated platelets (PubMed:11154118).
  • Target information above from: UniProt accession P09429 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt

Images

  • SDS-PAGE - Recombinant Human HMGB1 protein (ab167718)
    SDS-PAGE - Recombinant Human HMGB1 protein (ab167718)

    SDS-PAGE of reduced ab1677 stained with Coomassie Blue. 
    DTT-reduced Protein migrates as 28 kDa and 32 kDa due to different glycosylation.

  • Functional Studies - Recombinant Human HMGB1 protein (ab167718)
    Functional Studies - Recombinant Human HMGB1 protein (ab167718)

    ab167718 stimulates production of TNF-α in Raw-246.7 cells. The EC50 for this effect is 0.7855-0.8342 μg/mL.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (2)

Publishing research using ab167718? Please let us know so that we can cite the reference in this datasheet.

ab167718 has been referenced in 2 publications.

  • Shu Z  et al. The GSK-3ß/ß-catenin signaling pathway is involved in HMGB1-induced chondrocyte apoptosis and cartilage matrix degradation. Int J Mol Med 45:769-778 (2020). PubMed: 31922219
  • Lauenstein JU  et al. Negative Regulation of TLR Signaling by BCAP Requires Dimerization of Its DBB Domain. J Immunol 204:2269-2276 (2020). PubMed: 32198144

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Question

What are the recommended storage conditions

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Abcam community

Verified customer

Asked on Mar 08 2018

Answer

We strongly recommend reconstituting the protein as suggested and avoiding repeated freeze-thaw cycles. The reconstituted protein must NOT be aliquoted to less than 10 ug per vial to avoid surface adsorption loss, inactivation or shortening expiration date of protein.
We do know the reconstituted protein is stable within 3 months. Beyond 3 months, the protein may work well in your assay, but please kindly note that we could not provide warranty for it.
Please check whether adding the BSA or HSA in the protein would affect your assay. If no affect, it is recommended adding BSA or HSA for long term storage like more than 3 months.

Read More

Abcam Scientific Support

Answered on Mar 08 2018

This HMGB1 protein activates human TLR4

Excellent
Abreviews
Abreviews
abreview image
Application
Functional Studies
We tested this product's ability to stimulate human TLR4 using a reporter cell line from Invivogen (#hkb-htlr4). The cells were first treated with TLR4 inhibitor CLI-095 (Invivogen #tlrl-cli95) or vehicle (DMSO) for 6 hr, then exposed to lipopolysaccharide (Sigma L2762) or HMGB1 (Abcam ab167718). After 16 hr exposure, we assessed the levels of secreted embryonic alkaline phosphatase (proportional to amount of TLR4 activity in this cell line) by measuring optical density at 638 nm.

The result is a comparison of dose-response curves from LPS (the classic TLR4 agonist) and HMGB1. First, these results show that this product binds to and activates TLR4 (wells pre-treated with the TLR4 inhibitor CLI-095 showed no SEAP production data not shown). Second, it should be noted that this product is a less potent agonist than LPS, as evidenced by the curve being shifted to the right by about 2 orders of magnitude.

Abcam user community

Verified customer

Submitted Aug 03 2016

This HMGB1 induce inflammation in periodontal ligament fibroblast cell

Good
Abreviews
Abreviews
Application
ELISA
This HMGB1 promote the secretion of a concentration-dependent manner IL-8 in periodontal ligament fibroblast.

Abcam user community

Verified customer

Submitted Jul 22 2015

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