Rabbit HMGB1 Polyclonal Antibody | anti-HMGB1 antibody

HMGB1 Antibody

Cat. Num. AAA326757

• Gene Names: HMGB1; HMG1; HMG3; HMG-1; SBP-1
• Reactivity: Human, Mouse, Rat
• Applications: ELISA, Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Western Blot
• Purity: The antiserum was purified by peptide affinity chromatography using SulfoLink Coupling Resin.
• Synonyms: HMGB1, Antibody; HMGB1 Antibody; 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; anti-HMGB1 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat
• Clonality: Polyclonal
• Isotype: IgG
• Specificity: HMGB1 antibody detects endogenous levels of total HMGB1
• Purity/Purification: The antiserum was purified by peptide affinity chromatography using SulfoLink Coupling Resin.
• Form/Format: Liquid; Phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Concentration: 1mg/ml (varies by lot)
• Sequence Length: 215

• Applicable Applications for anti-HMGB1 antibody: ELISA, ICC (Immunocytochemistry), IF (Immunofluorescence), IHC (Immunohistochemistry), WB (Western Blot)
• Immunogen: A synthesized peptide derived from human HMGB1
• Subcellular Location: 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).
• Tissue Specificity: Ubiquituous. Expressed in platelets (PubMed:11154118).
• Conjugation: Unconjugated
• Preparation and Storage: Store at -20 degree C. Stable for 12 months from date of receipt.

IF (Immunofluorescence)

(AAA326757 staining Hela by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)

WB (Western Blot)

(Western blot analysis of extracts of Jurkat, using HMGB1antibody. The lane on the left is treated with the antigen-specific peptide.)

IF (Immunofluorescence)

(AAA326757 staining Hela by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)

WB (Western Blot)

(Western blot analysis of extracts of various tissue sample, using HMGB1 Antibody.)

IHC (Immunohistochemistry)

(AAA326757 at 1/100 staining Mouse kidney tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)

Related Product Information for anti-HMGB1 antibody

Description: High mobility group protein B1 (HMGB1) belongs to a family of highly conserved proteins that contain HMG box domains (1, 2). All three family members (HMGB1, HMGB2, and HMGB3) contain two HMG box domains and a C-terminal acidic domain. HMGB1 is a widely expressed and highly abundant protein (2). HMGB2 is widely expressed during embryonic development, but is restricted to lymphoid organs and testis in adult animals (3). HMGB3 is only expressed during embryogenesis (4). While expression varies, the biochemical properties of the different family members may be indistinguishable. The HMG box domains facilitate the binding of HMGB proteins to the minor groove of DNA, which results in local bending of the DNA double helix (1, 2). HMGB proteins are recruited by and help facilitate the assembly of site-specific DNA binding proteins to their cognate binding sites in chromatin. For example, HMGB1 facilitates the binding of Hox proteins, Oct-1, p53, Rel proteins, and steroid hormone receptor proteins to their target gene promoters (1, 2). In addition to their functions in the nucleus, HMGB proteins play a significant role in extracellular signaling associated with inflammation (5, 6). HMGB1 is massively released into the extracellular environment during cell necrosis, but not apoptosis. Extracellular HMGB1 alarms the innate immune system by acting as a chemoattractant for inflammatory leukocytes, smooth muscle cells, and stem cells, functioning as an immune adjuvant for soluble and particulate antigens, and triggering activation of T cells and dendritic cells. In addition, activated monocytes, macrophages and, dendritic cells also secrete HMGB1, forming a positive feedback loop that results in the release of additional cytokines and neutrophils. Hypoxia has also been shown to cause the release of HMGB1 in the liver, and some studies suggest a role for extracellular HMGB1 in tumor homeostasis (5, 6).
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 (PubMed:27362237). 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).
Subunit 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).
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 (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).
Similarity: 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). Belongs to the HMGB family.

References

Lu HY, Chen XQ, Tang W, Wang QX, Zhang J; Journal: Mol Med Rep. GRP78 silencing enhances hyperoxia-induced alveolar epithelial cell apoptosis via CHOP pathway.

NCBI and Uniprot Product Information

• NCBI GI #: 926459555
• NCBI GeneID: 3146
• NCBI Accession #: NP_001300821.1
• NCBI GenBank Nucleotide #: NM_001313892.1
• UniProt Accession #: P09429; Q14321; Q5T7C3; Q6IBE1; A5D8W9
• Molecular Weight: Observed: 25 kDa; Predicted: 25 kDa
• NCBI Official Full Name: high mobility group protein B1 isoform 1
• NCBI Official Synonym Full Names: high mobility group box 1
• NCBI Official Symbol: HMGB1
• NCBI Official Synonym Symbols: HMG1; HMG3; HMG-1; SBP-1
• NCBI Protein Information: high mobility group protein B1
• UniProt Protein Name: High mobility group protein B1
• UniProt Gene Name: HMGB1
• UniProt Synonym Gene Names: HMG1; HMG-1