HMGB1 Review 2017: HMGB1 antibodies and HMGB1 ELISA Kits

HMGB1 pedia

High mobility group box 1 (HMGB1) is a non-histone chromosome binding protein, and its function involves gene transcription, gene recombination, DNA damage repair and other biological processes. Overexpression of HMGB1 is one of the characteristics of certain tumors that correlates with the ability of tumors to proliferate, angiogenesis, and programmed death, and can attenuate the surveillance of growth signals, inflammatory responses, tumor tissue infiltration and metastasis effect. The study found that HMGB1 plays a central role in the development of tumors and the healing of multiple injuries. The role of HMGB1 in tumors, especially in the development of tumors, and the progress of targeted therapy targeting HMGB1 and its receptors have received extensive attention.

HMGB1 is present in almost all multicellular organisms. Initially, it was thought that it was rich in acidic amino acids and alkaline amino acids in a class of chromatin related proteins. HMGB1 is a highly conserved nuclear protein that binds to DNA as a chromosome binding protein and promotes transcription of related genes after protein synthesis. In the extracellular HMGB1 involved in inflammatory response, cell differentiation, cell migration and tumor metastasis. The interaction between HMGB1 and advanced glycation end product (RAGE) is closely related to the development of tumorigenesis.

HMGB1 Gene and Protein Databases

 

Alternative names for HMGB1 Gene

High Mobility Group Box 1, High-Mobility Group (Nonhistone Chromosomal) Protein 1, High Mobility Group Protein 1, Amphoterin, HMG-1, HMG1, High Mobility Group Protein B1, High-Mobility Group Box 1, SBP-1, HMG3

Databasae IDs

Human HMGB1 gene database

Entrez Gene Summary for HMGB1

This gene encodes 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.

GeneCards Summary for HMGB1

HMGB1 (High Mobility Group Box 1) is a Protein Coding gene. Diseases associated with HMGB1 include 13Q12.3 Microdeletion Syndrome and Adenosquamous Gallbladder Carcinoma. Among its related pathways are Activated TLR4 signalling and Immune System. GO annotations related to this gene include poly(A) RNA binding and chromatin binding. An important paralog of this gene is HMGB2.

Genomic Location for HMGB1 Gene

Chromosome:
13
Start:
30,456,704 bp from pter
End:
30,617,597 bp from pter
Size:
160,894 bases
Orientation:
Minus strand

HMGB1 Gene structure

A large number of sequences related to the cDNA for HMG1 protein hampered the cloning and mapping of the active HMG1 gene. Ferrari et al. (1996) showed that the HMG1 gene contains introns, whereas the HMG1-related sequences do not and most likely are retrotransposed pseudogenes. They identified 8 YACs from the ICI and CEPH libraries that contain the human HMG1 gene. The gene is similar in structure to the previously characterized mouse homolog.

Human HMGB1 protein database

HMGB1 protein attributes

Protein Symbol:
P09429-HMGB1_HUMAN
Recommended name:
High mobility group protein B1
Protein Accession:
P09429

Secondary Accessions:

  • A5D8W9
  • Q14321
  • Q5T7C3
  • Q6IBE1
Size:
215 amino acids
Molecular mass:
24894 Da
Quaternary structure:
  • Interacts (fully reduced HMGB1) with CXCL12; probably in a 1:2 ratio involving two molecules of CXCL12, each interacting with one HMG box of HMGB1; inhibited by glycyrrhizin (PubMed:22370717). Associates with the TLR4:LY96 receptor complex (PubMed:20547845). Component of the RAG complex composed of core components RAG1 and RAG2, and associated component HMGB1 or HMGB2 (By similarity). Interacts (in cytoplasm upon starvation) with BECN1; inhibits the interaction of BECN1 and BCL2 leading to promotion of autophagy (PubMed:20819940). Interacts with KPNA1; involved in nuclear import (PubMed:17114460). Interacts with SREBF1, TLR2, TLR4, TLR9, PTPRZ1, APEX1, FEN1, POLB, TERT (By similarity). Interacts with IL1B, AGER, MSH2, XPA, XPC, HNF1A, TP53 (PubMed:15014079, PubMed:18250463, PubMed:18160415, PubMed:19446504, PubMed:24474694, PubMed:23063560). Interacts with CD24; the probable CD24:SIGLEC10 complex is proposed to inhibit HGMB1-mediated tissue damage immune response (PubMed:19264983). Interacts with THBD; prevents HGMB1 interaction with ACER/RAGE and inhibits HGMB1 proinflammatory activity (PubMed:15841214). Interacts with HAVCR2; impairs HMGB1 binding to B-DNA and likely HMGB1-mediated innate immume response (By similarity). Interacts with XPO1; mediating nuclear export (By similarity). Interacts with HTT (wild-type and mutant HTT with expanded polyglutamine repeat) (PubMed:23303669).
Miscellaneous:
  • Proposed to contribute to the pathogenesis of various chronic inflammatory and autoimmune diseases, and cancer. High serum levels are found in several inflammatory events including sepsis, rheumatoid arthritis, artherosclerosis chronic kidney disease, systemic lupus erythematosus (SLE). Seems to be implicated in other diseases characterized by cell death and damage, including diabetes and Alzheimers disease. Its nucleosome-associated release during secondory necrosis may play a role in SLE (PubMed:19064698). During chemotherapy can mediate regrowth and metastasis of remaining cells in a AGER/RAGE-depenedent manner (PubMed:23040637). Purified HMG box 1 acts as a specific antogonist to HGMB1 pro-imflammatory activities (PubMed:14695889).

UniProtKB/Swiss-Prot for HMGB1

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).

HMGB1 protein sequence

 10 20 30 40 50 MGKGDPKKPR GKMSSYAFFV QTCREEHKKK HPDASVNFSE FSKKCSERWK 60 70 80 90 100 TMSAKEKGKF EDMAKADKAR YEREMKTYIP PKGETKKKFK DPNAPKRPPS 110 120 130 140 150 AFFLFCSEYR PKIKGEHPGL SIGDVAKKLG EMWNNTAADD KQPYEKKAAK 160 170 180 190 200 LKEKYEKDIA AYRAKGKPDA AKKGVVKAEK SKKKKEEEED EEDEEDEEEE 210 EDEEDEDEEE DDDDE 

Cloning and expression of HMGB1

The HMG (high mobility group) proteins are nonhistone chromosomal proteins. Wen et al. (1989) isolated a cDNA clone that encodes the entire 215-amino acid sequence of HMG1 from a human placenta cDNA library. Northern blot analysis showed that 3 mRNA species of approximately 1.0, 1.4, and 2.4 kb were expressed in all mammalian organs and cell lines examined. The HMG1 protein has a molecular mass of approximately 25 kD.

HMGB1 protein family and 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).
  • Belongs to the HMGB family.
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).
  • Contains 2 HMG box DNA-binding domains.
Family:
  • Belongs to the HMGB family.

HMGB1 protein Subcellular localization

Compartment Confidence
cytosol 5
extracellular 5
nucleus 5
plasma membrane 4
endosome 2
mitochondrion 2
cytoskeleton 1
endoplasmic reticulum 1
lysosome 1
vacuole 1

HMGB1 protein post-translational modifications

  • 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).
  • 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).
  • Phosphorylated at serine residues. Phosphorylation in both NLS regions is required for cytoplasmic translocation followed by secretion (PubMed:17114460).
  • Poly-ADP-ribosylated by PARP1 when secreted following stimulation with LPS (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).
  • Ubiquitination at Lys 43, Lys 112, and Lys 128
  • Modification sites at PhosphoSitePlus

HMGB1 related disease

Disorder Aliases PubMed IDs
13q12.3 microdeletion syndrome
 –
adenosquamous gallbladder carcinoma
 –
endophthalmitis  –
staphylococcal toxic shock syndrome
 –
anca-associated vasculitis

HMGB1 ELISA kit comparison

  • Human HMGB1 ELISA Kit (ARG81185)
  • $599
  • Arigo product: The best HMGB1 ELISA kit in Asian
  • Antibody: Monoclonal Only?
  • Protocol: 2 times incubation, 1 time rinse
  • (-60% time)

  • Sensitivity: 0.3ng/ml
  • (+233% sensitive)

  • Stability CV<5.2
  • (+92% stable)

  • Specificity: Cross-reaction with HMGB2 is < 2%?
  • Check now
  • The most popular HMGB1 Elisa kit
  • Price: Needs quote
  • The best Competitor Elisa Kit in the world
  • Antibody: Monoclonal & Polyclonal?
  • Protocol: 3 times incubation, 2 times rinse
  • (+40% time)

  • Sensitivity: 1ng/ml
  • (-70% sensitive)

  • Stability: CV<10
  • (-48% stable)

  • Specificity: Cross-reaction with HMGB2 is < 2%?
  • Oops!

Overview of ARG81185 Human HMGB1 ELISA Kit

Product Description ARG81185 Human HMGB1 ELISA Kit is an Enzyme Immunoassay kit in simple steps with good precision for the quantification of Human HMGB1 in plasma and cell culture supernatant.
Tested Reactivity Hu
Tested Application ELISA
Specificity This Human HMGB1 ELISA kit detect natural and recombinant Human HMGB1 protein and the cross-reactivity with HMGB2 is lower than <2%.
Target Name HMGB1
Sensitivity 0.3 ng/ml
Sample Type Plasma, and cell culture supernatant
Standard Range 0.3125 – 20 ng/ml
Sample Volume 100 µl
Precision < 5.2%
Full Name high mobility group box 1
Alternate Names HMG-1; High mobility group protein B1; High mobility group protein 1; HMG1; SBP-1; HMG3
Check now

HMGB1 antibodies recommendation

Name HMGB1 Polyclonal Antibody
Catalogue No. STJ96867
Reactivity Human, Mouse, Rat
Specificity HMGB1  Polyclonal Antibody detects endogenous levels of HMGB1  protein.
Immunogen Synthesized peptide derived from the N-terminal region of human HMGB1
Host Rabbit
Applications WB, IHC, ELISA
Recommended dilution WB 1:500-1:2000; IHC-P 1:100-1:300; ELISA 1:20000;
Clonality Polyclonal
Conjugation Unconjugated
Isotype IgG
Formulation HMGB1 Antibody was tube-contained. Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide.
Concentration 1 mg/ml
Purification HMGB1 Antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
Storage -20 Celsius degree. Avoid repeated freeze/thaw cycles.
Alternative antibody names High mobility group protein B1 antibody, High mobility group protein 1 antibody, HMGB1 antibody
Database links Human UniProt/Swiss-Prot:P09429;Mouse UniPort/Swiss-Prot: P63158;Rat UniProt/Swiss-Port: P63159;Human Entrez Gene: 3146;Mouse Entrez Gene: 15289;Rat Entrez Gene: Rn.144565
Protein names High mobility group protein B1 , High mobility group protein 1 , HMGB1
Check now