Instant ELISA Kit for High Mobility Group Protein 1 (HMGB1)

HMG1; HMG3; SBP1; Sulfoglucuronyl Carbohydrate Binding Protein; Amphoterin; High Mobility Group Box 1 Protein


This assay has high sensitivity and excellent specificity for detection of Instant High Mobility Group Protein 1 (HMGB1).
No significant cross-reactivity or interference between Instant High Mobility Group Protein 1 (HMGB1) and analogues was observed.


Matrices listed below were spiked with certain level of recombinant Instant High Mobility Group Protein 1 (HMGB1) and the recovery rates were calculated by comparing the measured value to the expected amount of Instant High Mobility Group Protein 1 (HMGB1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 96-103 99
EDTA plasma(n=5) 79-93 82
heparin plasma(n=5) 98-105 101


Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Instant High Mobility Group Protein 1 (HMGB1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Instant High Mobility Group Protein 1 (HMGB1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%


The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Instant High Mobility Group Protein 1 (HMGB1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 94-101% 79-91% 85-92% 83-96%
EDTA plasma(n=5) 83-90% 86-95% 81-99% 79-98%
heparin plasma(n=5) 89-98% 89-98% 96-105% 91-99%


The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.



Magazine Citations
Scandinavian Journal of Trauma, Resuscitation and Emergency Medicine High mobility group box protein-1 (HMGB-1) as a new diagnostic marker in patients with acute appendicitis PubMed: PMC3094252
Respiration Protective Effect of Nicotine on Lipopolysaccharide-Induced Acute Lung Injury in Mice Karger: 000319151
Surgery Clinical relevance of single nucleotide polymorphisms of the high mobility group box 1 protein gene in patients with major trauma in Southwest China Pubmed: 22047946
Viral Immunology Is HMGB1 a New Indirect Marker for Revealing Fibrosis in Chronic Hepatitis and a New Therapeutic Target in Treatment? Wiley: source
Chinese Medical Journal Severity of sepsis is correlated with the elevation of serum high-mobility group box 1 in rats CMJ: 2009218808935901080
Microbiology and Immunology Diagnostic value of serum concentrations of high-mobility group-box protein 1 and soluble hemoglobin scavenger receptor in brucellosis PubMed: 23278480
ACTA biochimica polonica High mobility group B1 levels in sepsis and disseminated intravascular coagulation Actabp: Source
Anesthesia and Analgesia The Inhibitory Effect of Lidocaine on the Release of High Mobility Group Box 1 in Lipopolysaccharide-Stimulated Macrophages PubMed: 21288974
Rheumatology International Elevated serum levels of high mobility group box protein 1 (HMGB1) in patients with ankylosing spondylitis and its association with disease activity and quality of life PubMed: 23143556
Inflammation Research Mammalian target of rapamycin complex 2 regulates inflammatory response to stress PubMed: 22899279
Acta Biochimica et Biophysica Sinica IGF-1 alleviates ox-LDL-induced inflammation via reducing HMGB1 release in HAECs PubMed: 22782142
LabVest The suppression of TRIM21 and the accumulation of IFN-α play crucial roles in the pathogenesis of osteonecrosis of the femoral head. PubMed: 22825687
Biomark Insights Decreased Epidermal Growth Factor (EGF) Associated with HMGB1 and Increased Hyperactivity in Children with Autism PubMed: PMC3623607
Evidence-based Complementary and Alternative Medicine ATF3 Protects against LPS-Induced Inflammation in Mice via Inhibiting HMGB1 Expression PubMed: PMC3770047
Exp Ther Med Correlation between serum high-mobility group box-1 levels and high-sensitivity C-reactive protein and troponin I in patients with coronary artery disease Pubmed: 23935732
International of Rheumatic disesases Serum levels of high mobility group box 1 protein and its association with quality of life and psychological and functional status in patients with fibromyalgia Pubmed: 23901875
Surgery A novel serum marker for biliary tract cancer: Diagnostic and prognostic values of quantitative evaluation of serum mucin 5AC (MUC5AC) Pubmed: 24468034
Acta Pharmacol Sin. Simvastatin suppresses vascular inflammation and atherosclerosis in ApoE-/- mice by downregulating the HMGB1-RAGE axis Pubmed: 23564080
BMC pulmonary medicine Soluble receptor for advanced glycation end-products and progression of airway disease Pubmed: 24758342
Biochemical Pharmacology Ascorbic acid reduces HMGB1 secretion in lipopolysaccharide-activated RAW 264.7 cells and improves survival rate in septic mice by activation of Nrf2/HO-1 signals Pubmed:25896849
BioMed Research International Effects of Single or Combined Treatments with Radiation and Chemotherapy on Survival and Danger Signals Expression in Glioblastoma Cell Lines Hindawi:Source
J Cardiothorac Surg. High plasma levels of high mobility group box 1 is associated with the risk of sepsis in severe blunt chest trauma patients: a prospective cohort study Pubmed:Pmc4132233
J Leukoc Biol. HMGB1 is an early and critical mediator in an animal model of uveitis induced by IRBP-specific T cells Pubmed:24374967
PLOS ONE The Alarmin Concept Applied to Human Renal Transplantation: Evidence for a Differential Implication of HMGB1 and IL-33 Pubmed:24586382
Journal of Surgical Research Plasma levels of high mobility group box 1 increase in patients with posttraumatic stress disorder after severe blunt chest trauma: a prospective cohort study Pubmed:25016440
Virus Research High-mobility group box-1, promising serological biomarker for the distinction of human WNV disease severity Pubmed:25197038
Journal of Stroke and Cerebrovascular Diseases Increased VEGF and Decreased SDF-1α in Patients with Silent Brain Infarction Are Associated with Better Prognosis after First-Ever Acute Lacunar Stroke Pubmed:25601176
中华临床医师杂志 高迁移率族蛋白B1和神经元烯醇化酶对心脏骤停大鼠复苏后脑损伤评价的研究 Journal: Fulltext
Oxidative Medicine and Cellular Longevity Diabetes-Induced Oxidative Stress in Endothelial Progenitor Cells May Be Sustained by a Positive Feedback Loop Involving High Mobility Group Box-1 Journals: Omcl
Int Immunopharmacol Hesperidin ameliorates lipopolysaccharide-induced acute lung injury in mice by inhibiting HMGB1 release PubMed: 25724384
Journal of Immunotherapy Wogonin Inhibits Tumor-derived Regulatory Molecules by Suppressing STAT3 Signaling to Promote Tumor Immunity PubMed: 25962106
Atherosclerosis Retinoic acid inhibits tissue factor and HMGB1 via modulation of AMPK activity in TNF-α activated endothelial cells and LPS-injected mice PubMed: 26116962
PLOS ONE Plasma HMGB-1 Levels in Subjects with Obesity and Type 2 Diabetes: A Cross-Sectional Study in China PubMed: 26317615
Int Immunopharmacol Betulin attenuates lung and liver injuries in sepsis PubMed: 26644168
journal of cardiochoracic surgery High mobility group box 1 gene polymorphism is associated with the risk of postoperative atrial fibrillation after coronary artery bypass surgery PubMed: 26109393
Int Immunopharmacol Glycyrrhizin reduces HMGB1 secretion in lipopolysaccharide-activated RAW 264.7 cells and endotoxemic mice by p38/Nrf2-dependent induction of HO-1 PubMed: 25812767
World J Gastroenterol Overexpression of HMGB1 A-box reduced lipopolysaccharide-induced intestinal inflammation via HMGB1/TLR4 signaling in vitro PubMed: 26167076
Neurotoxicology Allopurinol reduces severity of delayed neurologic sequelae in experimental carbon monoxide toxicity in rats PubMed: 25845300
Journal of Neuroimmunology HMGB1 expression patterns during the progression of experimental autoimmune encephalomyelitis PubMed: 25773152
J Neuroinflammation HMGB1 release triggered by the interaction of live retinal cells and uveitogenic T cells is Fas/FasL activation-dependent PubMed: 26394985
PLoS One Low Shear Stress Induced HMGB1 Translocation and Release via PECAM-1/PARP-1 Pathway to Induce Inflammation Response PubMed: 25793984
Neuroscience Interleukin-33 is released in spinal cord and suppresses experimental autoimmune encephalomyelitis in mice PubMed: 26363151
Cancer Biomarkers Serum levels of HMGB1 have a diagnostic role in metastatic renal cell cancer Pubmed:27062570
International Immunopharmacology Oxymatrine attenuates CCl 4-induced hepatic fibrosis via modulation of TLR4-dependent inflammatory and TGF-β1 signaling pathways Pubmed:27179304
SpringerPlus Ethyl pyruvate attenuated coxsackievirus B3-induced acute viral myocarditis by suppression ofHMGB1/RAGE/NF-ΚB pathway. Pubmed:27026909
Acta Biochim Biophys Sin (Shanghai). High mobility group box1 protein is involved in acute inflammation induced by Clostridium difficile toxin A. content:2016
United States Patent Application METHOD OF MAKING A SKIN CARE COMPOSITION y2016:0077080.html
Oxid Med Cell Longev. Oxidative Stress in Metabolic Disorders: Pathogenesis, Prevention, and Therapeutics Pubmed:26949450
Oncotarget Zoledronic acid-encapsulating self-assembling nanoparticles and doxorubicin: a combinatorial approach to overcome simultaneously chemoresistance and immunoresistance in breast tumors pubmed:26980746
clinical biochemistry Relevance of pre-analytical blood management on the emerging cardiovascular protein biomarkers TWEAK and HMGB1 and on miRNA serum and plasma profiling. pubmed:27847340
disease markers Serum HMGB1 as a Potential Biomarker for Patients with Asbestos-Related Diseases. pubmed:28348451
Biogerontology. Changes in pro-inflammatory markers and leucine concentrations in response to Nordic Walking training combined with vitamin D supplementation in elderly women. pubmed:28316011
Journal of Immunology Research Serum HMGB1 serves as a novel laboratory indicator reflecting disease activity and pubmed:27800496
Springerplus Ethyl pyruvate attenuated coxsackievirus B3-induced acute viral myocarditis by suppression of HMGB1/RAGE/NF-ΚB pathway pubmed:27026909
Molecular Immunology HMGB1 blockade differentially impacts pulmonary inflammation and defense responses in poly (I: C)/LPS-exposed heart transplant mice pubmed:27387278
Oxidative Medicine and Cellular Longevity Reduced HMGB 1-Mediated Pathway and Oxidative Stress in Resveratrol-Treated Diabetic Mice: A Possible Mechanism of Cardioprotection of Resveratrol in … pubmed:27833703
BioMed Research International  High Mobility Group Box1 Protein Is Involved in Endoplasmic Reticulum Stress Induced by Clostridium difficile Toxin A pubmed:27579314
Cardiology The role of high mobility group box 1 protein in interleukin-18-induced myofibroblastic transition of valvular interstitial cells pubmed:27395056
Innate immunity HMGB1 exacerbates experimental mouse colitis by enhancing innate lymphoid cells 3 inflammatory responses via promoted IL-23 production pubmed:27670944
Urolithiasis. Urinary MCP-1、HMGB1 increased in calcium nephrolithiasis patients and the influence ofhypercalciuria on the production of the two cytokines. pubmed:27393275
Journal of Investigative Surgery Diagnostic Value of High-Mobility Group Box 1 (HMGB1) Protein in Acute and Perforated Appendicitis. pubmed:27922766
J Cell Biochem High Mobility Group Box 1 Mediates Interferon-r-Induced Phenotypic Modulation of Vascular Smooth Muscle Cells. pubmed:27579780
BMC Vet Res. Blood markers of fibrinolysis and endothelial activation in canine babesiosis. pubmed:28363279
Immunol Res. Glycyrrhizin ameliorates experimental colitis through attenuating interleukin-17-producing T cell responses via regulating antigen-presenting cells. pubmed:28108937
Biomed Pharmacother. SIRT1 inhibits releases of HMGB1 and HSP70 from human umbilical vein endothelial cells caused by IL-6 and the serum from a preeclampsia patient and protects the cells from death. pubmed:28122311
Sci Rep.  Bone marrow-derived mesenchymal stem cells (BMSCs) repair acute necrotized pancreatitis by secreting microRNA-9 to target the NF-κB1/p50 gene in rats. pubmed:28373667
Integrative Cancer Science and Therapeutics The expression of HMGB1 and the receptor for advanced glycation end products (RAGE) in epithelial ovarian cancer pdf:ICST-4-232.pdf
Journal of Food and Drug Analysis Anti-inflammatory effect of cinnamaldehyde and linalool from the leaf essential oil of Cinnamomum osmophloeum Kanehira in endotoxin-induced … S1021-9498(17)30081-9
Biomedicine & Pharmacotherapy SIRT1 inhibits releases of HMGB1 and HSP70 from human umbilical vein endothelial cells caused by IL-6 and the serum from a preeclampsia patient and protects the cells from death DOI:10.1016/j.biopha.2017.01.087
Stem Cell Rev and Rep Induction of Immunogenic Cell Death in Lymphoma Cells by Wharton’s Jelly Mesenchymal Stem Cell Conditioned Medium pubmed:28918474
Scientific Reports COMMUNICATIONS Mineral particles stimulate innate immunity through neutrophil extracellular traps containing HMGB1 pubmed:29192209
Biochemical and Biophysical Research Communications Oxidized low-density lipoprotein promotes osteoclast differentiation from CD68 positive mononuclear cells by regulating HMGB1 release pubmed:29146189
Pulmonary Pharmacology & Therapeutics HMGB1 promotes HLF-1 proliferation and ECM production through activating HIF1-a-regulated aerobic glycolysis pubmed:28571757
Biomedical Research Correlation between high mobility group box 1 protein, tumor necrosis factor-α, and acute coronary syndrome ISSN 0970-938X
Molecular Cancer PERK induces resistance to cell death elicited by endoplasmic reticulum stress and chemotherapy pubmed:28499449
BMC Complementary and Alternative Medicine Assessing the recovery from prerenal and renal acute kidney injury after treatment with single herbal medicine via activity of the biomarkers HMGB1, NGAL and KIM-1 in kidney proximal tubular cells treated by cisplatin with different doses and exposure times. pubmed:29258482
25 Blocking the Feedback Loop between Neuroendocrine Differentiation and Macrophages Improves the Therapeutic Effects of Enzalutamide (MDV3100) on Prostate Cancer. pubmed:29191973
Nature microbiology Secreted tryptophanyl-tRNA synthetase as a primary defence system against infection. pubmed:27748732
Cellular physiology and biochemistry Macrophage Inflammatory Protein-2 in High Mobility Group Box 1 Secretion of Macrophage Cells Exposed to Lipopolysaccharide. pubmed:28662496
Molecular Medicine Reports Niclosamide attenuates inflammatory cytokines via the autophagy pathway leading to improved outcomes in renal ischemia/reperfusion injury pubmed:28627643
Cell Death & Disease AMP-activated protein kinase agonist N 6-(3-hydroxyphenyl) adenosine protects against fulminant hepatitis by suppressing inflammation and apoptosis Articles:s41419-017-0118-0
Journal of Molecular Cell Biology S1 PR 3 is essential for phosphorylated fingolimod to protect astrocytes against oxygen‐glucose deprivation‐induced neuroinflammation via inhibiting TLR 2/4‐NF κB … Pubmed:29536648
Frontiers in Immunology Glycyrrhizin Protects Mice against Experimental Autoimmune Encephalomyelitis by Inhibiting HMGB1 Expression and Neuronal HMGB1 Release Pubmed:30013568
Oncoimmunology Bromodomain inhibition exerts its therapeutic potential in malignant pleural mesothelioma by promoting immunogenic cell death and changing the tumor immune … Pubmed:29399399
Cancer Immunology, Immunotherapy Tumor lysate-based vaccines: on the road to immunotherapy for gallbladder cancer Pubmed:29600445
Cytokine Porcine reproductive and respiratory syndrome virus induces concurrent elevation of High Mobility Group Box-1 protein and pro-inflammatory cytokines in … Pubmed:29895396
Frontiers in Neurology Ursolic Acid Ameliorates Inflammation in Cerebral Ischemia and Reperfusion Injury Possibly via High Mobility Group Box 1/Toll-Like Receptor 4/NFκB … Pubmed:29867706
Biomedicine & Pharmacotherapy Oridonin enhances the cytotoxicity of 5-FU in renal carcinoma cells by inducting necroptotic death Pubmed:29958141
Inflammation Research Luteolin activates ERK1/2- and Ca-dependent HO-1 induction that reduces LPS-induced HMGB1, iNOS/NO, and COX-2 expression in RAW264.7 cells and … Pubmed:29497773
Journal of Food and Drug Analysis Anti-inflammatory effect of cinnamaldehyde and linalool from the leaf essential oil of Cinnamomum osmophloeum Kanehira in endotoxin-induced mice Pubmed:29389558
Biochemical and Biophysical Research Communications Sirt1 S-nitrosylation induces acetylation of HMGB1 in LPS-activated RAW264. 7 cells and endotoxemic mice Pubmed:29680657
Neurological Sciences Thrombomodulin, alarmin signaling, and copeptin: cross-talk between obesity and acute ischemic stroke initiation and severity in Egyptians Pubmed:29637447
International Journal of Gynecology & Obstetrics Increased serum levels of high mobility group protein B1 and calprotectin in pre‐eclampsia Pubmed:29569400
Advances in Experimental Medicine and Biology Molecules of Damage-Associated Patterns in Bronchoalveolar Lavage Fluid and Serum in Chronic Obstructive Pulmonary Disease Pubmed:29429028
Taiwanese Journal of Obstetrics & Gynecology HMGB1-RAGE signaling pathway in pPROM Pubmed:29673663
Chronic obstructive pulmonary disease (COPD)--from biomarkers to clinical phenotypes ISBN:978-951-51-4039-5
Journal of Pharmacological Sciences Butyrate suppresses abnormal proliferation in colonic epithelial cells under diabetic state by targeting HMGB1 Doi: 10.1016/j.jphs.2018.07.012
Biomaterials Science Delivery of 5'-Triphosphate RNA with Endosomolytic Nanoparticles Potently Activates RIG-I to Improve Cancer Immunotherapy Doi: 10.1039/C8BM01064A 
Toxicological Sciences Apoptosis resistance in fibroblasts precedes progressive scarring in pulmonary fibrosis and is partially mediated by Toll-like receptor 4 activation Pubmed: 31020321
Molecular Medicine FBXW7 suppresses HMGB1-mediated innate immune signaling to attenuate hepatic inflammation and insulin resistance in a mouse model of nonalcoholic … Pubmed: 31215394
international immunopharmacology Dulaglutide mitigates inflammatory response in fibroblast-like synoviocytes Pubmed: 31185450
Food & Function Network pharmacology oriented study reveals inflammatory state-dependent dietary supplement hepatotoxicity responses in normal and diseased rats Pubmed: 31140472
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