• Human HMGB1 (High mobility group protein B1) CLIA Kit (STJC0000695)

Human HMGB1 (High mobility group protein B1) CLIA Kit (STJC0000695)

SKU:
STJC0000695

Shipping:
Free Shipping
Current Stock:
Applications: CLIA
Reactivity: Human
Note: FOR SCIENTIFIC EDUCATIONAL RESEARCH USE ONLY (RUO). MUST NOT BE USED IN DIAGNOSTIC OR OTHER MEDICAL APPLICATIONS.
Sensitivity: 7.5pg/mL
Detection Limit: 12.5~800pg/mL
Short Description: This human HMGB1 kit is a highly sensitive in-vitro chemiluminescent immunoassay for the measurement of trace amounts of analytes.
Storage Instruction: If unopened the kit may be stored at 2-8°C for up to 1 month. If the kit will not be used within 1 month, store the components separately, according to the component table in the manual.
Assay Time: 3.5h
Detection: Chemiluminescence
Gene Symbol: HMGB1
Gene ID: 3146
Uniprot ID: HMGB1_HUMAN
Specificity: This kit recognizes Human HMGB-1 in samples. No significant cross-reactivity or interference between Human HMGB-1 and analogues was observed.
Sample Type: Serum, plasma and other biological fluids
Tissue Specificity Ubiquitous. Expressed in platelets.
Post Translational Modifications Phosphorylated at serine residues. Phosphorylation in both NLS regions is required for cytoplasmic translocation followed by secretion. Acetylated on multiple sites upon stimulation with LPS. Acetylation on lysine residues in the nuclear localization signals (NLS 1 and NLS 2) leads to cytoplasmic localization and subsequent secretion. 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. Can be proteolytically cleaved by a thrombin:thrombomodulin complex.reduces binding to heparin and pro-inflammatory activities. Forms covalent cross-links mediated by transglutaminase TGM2, between a glutamine and the epsilon-amino group of a lysine residue, forming homopolymers and heteropolymers.
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. Has proangiogdenic activity. May be involved in platelet activation. Binds to phosphatidylserine and phosphatidylethanolamide. Bound to RAGE mediates signaling for neuronal outgrowth. May play a role in accumulation of expanded polyglutamine (polyQ) proteins such as huntingtin (HTT) or TBP. 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. May have an enhancing role in nucleotide excision repair (NER). However, effects in NER using in vitro systems have been reported conflictingly. May be involved in mismatch repair (MMR) and base excision repair (BER) pathways. May be involved in double strand break repair such as non-homologous end joining (NHEJ). 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). In vitro can displace histone H1 from highly bent DNA. Can restructure the canonical nucleosome leading to relaxation of structural constraints for transcription factor-binding. Enhances binding of sterol regulatory element-binding proteins (SREBPs) such as SREBF1 to their cognate DNA sequences and increases their transcriptional activities. Facilitates binding of TP53 to DNA. Proposed to be involved in mitochondrial quality control and autophagy in a transcription-dependent fashion implicating HSPB1.however, this function has been questioned. 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. Involved in oxidative stress-mediated autophagy. 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. In myeloid cells has a protective role against endotoxemia and bacterial infection by promoting autophagy. 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. Induces the migration of monocyte-derived immature dendritic cells and seems to regulate adhesive and migratory functions of neutrophils implicating AGER/RAGE and ITGAM. 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. Promotes extracellular DNA-induced AIM2 inflammasome activation implicating AGER/RAGE. 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. Promotes secretion of interferon-gamma by macrophage-stimulated natural killer (NK) cells in concert with other cytokines like IL-2 or IL-12. TLR4 is proposed to be the primary receptor promoting macrophage activation and signaling through TLR4 seems to implicate LY96/MD-2. 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. Contributes to tumor proliferation by association with ACER/RAGE. Can bind to IL1-beta and signals through the IL1R1:IL1RAP receptor complex. 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. 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. In adaptive immunity may be involved in enhancing immunity through activation of effector T cells and suppression of regulatory T (TReg) cells. 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. Also reported to limit proliferation of T-cells. Released HMGB1:nucleosome complexes formed during apoptosis can signal through TLR2 to induce cytokine production. 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. During macrophage activation by activated lymphocyte-derived self apoptotic DNA (ALD-DNA) promotes recruitment of ALD-DNA to endosomes. (Microbial infection) Critical for entry of human coronaviruses SARS-CoV and SARS-CoV-2, as well as human coronavirus NL63/HCoV-NL63. Regulates the expression of the pro-viral genes ACE2 and CTSL through chromatin modulation. Required for SARS-CoV-2 ORF3A-induced reticulophagy which induces endoplasmic reticulum stress and inflammatory responses and facilitates viral infection. (Microbial infection) Associates with the influenza A viral protein NP in the nucleus of infected cells, promoting viral growth and enhancing the activity of the viral polymerase. (Microbial infection) Promotes Epstein-Barr virus (EBV) latent-to-lytic switch by sustaining the expression of the viral transcription factor BZLF1 that acts as a molecular switch to induce the transition from the latent to the lytic or productive phase of the virus cycle. Mechanistically, participates in EBV reactivation through the NLRP3 inflammasome. (Microbial infection) Facilitates dengue virus propagation via interaction with the untranslated regions of viral genome. In turn, this interaction with viral RNA may regulate secondary structure of dengue RNA thus facilitating its recognition by the replication complex.
Protein Name High Mobility Group Protein B1
High Mobility Group Protein 1
Hmg-1
Database Links Reactome: R-HSA-1236974
Reactome: R-HSA-140342
Reactome: R-HSA-166058
Reactome: R-HSA-445989
Reactome: R-HSA-5602498
Reactome: R-HSA-5603041
Reactome: R-HSA-5620971
Reactome: R-HSA-5686938
Reactome: R-HSA-6798695
Reactome: R-HSA-879415
Reactome: R-HSA-933542
Cellular Localisation Nucleus
Chromosome
Cytoplasm
Secreted
Cell Membrane
Peripheral Membrane Protein
Extracellular Side
Endosome
Endoplasmic Reticulum-Golgi Intermediate Compartment
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
Passively Released To The Extracellular Milieu From Necrotic Cells By Diffusion
Involving The Fully Reduced Hgmb1 Which Subsequently Gets Oxidized
Also Released From Apoptotic Cells
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
Secreted By Plasma Cells In Response To Lps
Found On The Surface Of Activated Platelets
An Increased Chromatin Association Is Observed When Associated With The Adenovirus Protein Pvii
Endoplasmic Reticulum
(Microbial Infection) Sars-Cov-2 Orf3a Promotes Hmgb1 Translocation From The Nucleus To The Cytoplasm Where It Is Recruited By And Colocalizes With Orf3a At The Endoplasmic Reticulum
Alternative CLIA Names High Mobility Group Protein B1 CLIA kit
High Mobility Group Protein 1 CLIA kit
Hmg-1 CLIA kit
HMGB1 CLIA kit
HMG1 CLIA kit
Specificity This kit recognizes Human HMGB-1 in samples. No significant cross-reactivity or interference between Human HMGB-1 and analogues was observed.
Reproducibility Both intra-CV and inter-CV are

Information sourced from Uniprot.org


Item Specifications Storage
Micro CLIA Plate (Dismountable) 96T: 8 wells ×12 strips strips -20℃, 6 months
Reference Standard 96T: 2 vials 48T: 1 vial -20℃, 6 months
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL 60 μL -20℃, 6 months
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL 60 μL -20℃ (Protect from light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL 2-8°C, 6 months
HRP Conjugate Diluent 1 vial, 14 mL 2-8°C, 6 months
Concentrated Wash Buffer (25×) 1 vial, 30 mL 2-8°C, 6 months
Substrate Reagent A 1 vial, 5 mL 2-8℃ (Protect from light)
Substrate Reagent B 1 vial, 5 mL 2-8℃ (Protect from light)
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Sample Type Range (%) Average Recovery (%)
Serum(n=8) 99-111 104
EDTA plasma(n=8) 94-111 102
Cell culture media(n=8) 86-99 91

Intra-assay Precision Intra-assay Precision Intra-assay Precision Inter-assay Precision Inter-assay Precision Inter-assay Precision
Sample 1.00 2.00 3.00 1.00 2.00 3.00
n 20.00 20.00 20.00 20.00 20.00 20.00
Mean (pg/mL) 38.94 71.20 397.79 37.00 72.83 379.26
Standard deviation 4.74 8.16 27.92 3.48 5.48 28.71
CV (%) 12.17 11.46 7.02 9.41 7.52 7.57
12 months for antibodies. 6 months for ELISA Kits. Please see website T&Cs for further guidance