Human High mobility group protein B1,HMGB-1 ELISA Kit

Instructions
Code CSB-E08223h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human High mobility group protein B1 (HMGB1) ELISA Kit allows for the in vitro quantitative determination of HMGB1 concentrations in serum, plasma, tissue homogenates, or cell lysates. This kit exclusively recognizes human HMGB1. HMGB1 is a nonhistone nucleoprotein and also an extracellular inflammatory cytokine. Extracellular HMGB1 acting as a damage-associated molecular pattern (DAMP) binds with its pattern recognition receptors (PRRs), mediating the proliferation, differentiation, mobilization, and senescence of hematopoietic stem cells (HSCs). HMGB1 also participates in the formation of the inflammatory bone marrow (BM) microenvironment by activating proinflammatory signaling pathways. It plays an important role in the pathogenesis of hematopoietic malignancies. Extensively DNA-bound intracellular HMGB1 is involved in transcriptional modulation, DNA replication and repair, telomere maintenance, and nucleosome assembly.

The detection mechanism of this kit is based on the Sandwich-ELISA technique. HMGB1 in the sample is bound to the capture antibody immobilized on the microtiter plate and then sandwiched with the Biotin-labeled HMGB1 antibody. The solution color develops into blue after the ordinal addition of HRP-avidin and TMB. The color development is terminated after adding the stop solution, and the color turns from blue to yellow. The color intensity is positively correlated with HMGB1 content in the sample. The kit has been quality controlled with high sensitivity, strong specificity, good linearity, precision less than 10%, recovery with 85%-102%, and high lot-to-lot consistency. Refer to the product manual for more information.

Target Name high-mobility group box 1
Alternative Names Amphoterin ELISA Kit; Chromosomal protein; nonhistone; HMG1 ELISA Kit; DKFZp686A04236 ELISA Kit; High mobility group 1 ELISA Kit; High mobility group box 1 ELISA Kit; High mobility group protein 1 ELISA Kit; High mobility group protein B1 ELISA Kit; high-mobility group (nonhistone chromosomal) protein 1 ELISA Kit; HMG-1 ELISA Kit; HMG1 ELISA Kit; HMG3 ELISA Kit; HMGB 1 ELISA Kit; HMGB1 ELISA Kit; HMGB1_HUMAN ELISA Kit; NONHISTONE CHROMOSOMAL PROTEIN HMG1 ELISA Kit; SBP 1 ELISA Kit; Sulfoglucuronyl carbohydrate binding protein ELISA Kit
Abbreviation HMGB1
Uniprot No. P09429
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates, cell lysates
Detection Range 78 pg/mL-5000 pg/mL
Sensitivity 19.5 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Epigenetics and Nuclear Signaling
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%        
Three samples of known concentration were tested twenty times on one plate to assess.    
Inter-assay Precision (Precision between assays): CV%<10%        
Three samples of known concentration were tested in twenty assays to assess.      
               
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human HMGB-1 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.  
  Sample Serum(n=4)    
1:1 Average % 90    
Range % 84-99    
1:2 Average % 88    
Range % 82-94    
1:4 Average % 95    
Range % 89-102    
1:8 Average % 94    
Range % 88-100    
Recovery
The recovery of human HMGB-1 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.  
 
Sample Type Average % Recovery Range    
Serum (n=5) 98 90-102    
EDTA plasma (n=4) 91 85-100    
               
               
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.  
 
pg/ml OD1 OD2 Average Corrected    
5000 1.912 1.831 1.872 1.780    
2500 1.454 1.369 1.412 1.320    
1250 0.839 0.789 0.814 0.722    
625 0.456 0.487 0.472 0.380    
312 0.283 0.297 0.290 0.198    
156 0.199 0.202 0.201 0.109    
78 0.103 0.108 0.106 0.014    
0 0.089 0.094 0.092      
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with anti-human HMGB1 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard --- Dilute a bottle of standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled HMGB1 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the high concentration Biotin-antibody to an appropriate working solution.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the high concentration HRP-avidin solution to an appropriate solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) --- Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) --- Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator that can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

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Target Data

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
Gene References into Functions
  1. Based on these findings, ischemia/reperfusion-induced MCPIP1 expression regulates the migration and apoptosis of human vascular endothelial cells via HMGB1 and CaSR, respectively. PMID: 29379093
  2. Studied association of plasma levels of high mobility group box 1 (HMGB1) in critically ill patients. PMID: 29862569
  3. The high-mobility group box (HMGB) proteins, particularly HMGB1, are self-derived innate immune activators that have multiple functions in the regulation of immunity and inflammation. Recent discoveries have illustrated the close link between HMGB1 and heart allograft rejection. PMID: 29198620
  4. miR-193a plays a suppressive role in osteogenic differentiation of human bone marrow-derived stroma cell via targeting HMGB1. PMID: 29787753
  5. Platelet HMGB1 mediated neutrophil-extracellular traps release is a primary regulator of deep vein thrombosis in mice. PMID: 29391442
  6. Study findings suggested that TCTP promotes colorectal cancer metastasis through regulating the behaviors of HMGB1 and the downstream activation of the NF-kappaB signaling pathway. PMID: 30066846
  7. in the present study we explore the characteristic binding mode and energetics of Act D binding to 21 nt mimed CpG sequence in the positive regulatory region of hmgb1 gene to identify areas of pressing experimental need.. PMID: 28033959
  8. Study demonstrated that serum HMGB1 was upregulated and the expression levels of miR-381 were downregulated in patients with polymyositis (PM). Furthermore, high HMGB1 expression was associated with poor survival rate in patients with PM. A luciferase activity assay was used to confirm the binding of miR-381 and HMGB1 3' untranslated region. PMID: 29956737
  9. High mobility group box 1 (HMGB1) and OV-6 antigen (OV-6) positive staining are promising prognostic parameters for hepatocellular carcinoma (HCC), suggesting that HMGB1 and OV-6 may cooperate with each other and predict poor prognosis of HCC. PMID: 29441453
  10. results demonstrate that non-oxidizable HMGB1 induce a sustained cardiac fibroblasts migration despite the redox state of the environment and by altering CXCL12/CXCR4 axis. This affects proper cardiac remodeling after an infarction. PMID: 28716707
  11. High HMGB1 expression is associated with reduced chemosensitivity in the pleural effusion of non-small cell lung cancer. PMID: 28885675
  12. This work indicates that NELL-1, HMGB1, and CCN2 might enhance bone defect healing via the recruitment of endogenous cells and induction of vascularization and act via different processes than BMP2. PMID: 28463604
  13. comparison of inhibitory potential of the already known inhibitors of Head and neck squamous cell carcinoma against HMGB1-binding pocket using simulations and docking. PMID: 27900730
  14. identified 2 novel miR-193a-3p targets, the high mobility group box-1 (HMGB1) and the hypoxia upregulated-1 (HYOU1) gene products. HMGB1 silencing in cord blood ECFC-derived cells confirmed its role in regulating vascular function. PMID: 28276476
  15. Results show that HMGB1 is highly expressed in prostate cancer (PC) tissues. Highly expressed HMGB1 activates RAGE/NF-kappaB signaling pathways, which facilitates the metastasis of prostate cancer. PMID: 29845254
  16. HMGB1 was identified as a down-stream target of miR-204. The miR-204/HMGB1 axis mediated ZEB2-AS1's effect on pancreatic cancer. PMID: 29753015
  17. Study demonstrated that HMGB1 and TLR4 could contribute to the inflammatory lichen planus process in skin. PMID: 29728859
  18. Our research shows that HMGB1 participates in autophagy and DNA damage repair and that downregulation of HMGB1 enhances the sensitivity of multiple myeloma (MM)cells to Dex, suggesting that HMGB1 may serve as a target for MM treatment. PMID: 30157958
  19. HMGB1 plays a critical role in mitochondrial autophagy in cardiomyocytes. miR-410 targets its 3'UTR and regulates its transcription. PMID: 28914970
  20. HMGB1 is a good diagnostic biomarker for differentiating refractory M. pneumoniae pneumonia and non-refractory M. pneumoniae pneumonia. PMID: 30157804
  21. HMGB1 promoted lung cancer invasion and metastasis by upregulating the expression and activity of MMP-2 in an NF-kappaB-dependent manner. PMID: 29850505
  22. meta-analysis exploring the association of four HMGB1 polymorphisms with cancer. PMID: 29730397
  23. levels of serum HMGB1 were positively associated with 10-year CHD risk PMID: 29704473
  24. high mobility group box 1-receptor for advanced glycation end-products (HMGB1-RAGE) signaling pathway may be involved in the pathogenesis of preterm premature rupture of membranes (pPROM) PMID: 29673663
  25. MiR-106 interacted with the 3'-UTR of HMGB1 and inhibited HMGB1 expression. PMID: 30055307
  26. It is a 30 kDa protein that is a lethal mediator in sepsis and is a recognized therapeutic target. PMID: 30135341
  27. HMGB1 knockdown and miR-505 overexpression promoted ADM-induced DNA damage in HCC cells. PMID: 29803174
  28. These findings indicated that HMGB1 likely regulates autophagy in LO2 cells exposed to anoxia-reoxygenation injury PMID: 29880383
  29. extracellular functions of HMGB1 in cellular and immune homeostasis at the airway mucosal surface PMID: 28976774
  30. Our results indicate that genetic variations in the HMGB1 gene may serve as an important predictor of breast cancer progression and metastasis. PMID: 29725248
  31. soluble CD52 exerts a concerted immunosuppressive effect by first sequestering HMGB1 to nullify its proinflammatory Box B, followed by binding to the inhibitory Siglec-10 receptor, triggering recruitment of SHP1 to the intracellular immunoreceptor tyrosine-based inhibitory motif of Siglec-10 and its interaction with the TCR. PMID: 29997173
  32. In lung cancer patients, circulating HMGB1-containing nucleosome is higher in those under chemotherapy, predicting poorly cancer cell differentiation state, enhanced cancer invasion and advanced TNM stages. PMID: 29679570
  33. a number of microRNAs (miRNAs) are identified as a class of regulators for broad control of HMGB1-mediated biological actions in eukaryotic cells. PMID: 29651425
  34. The study indicated polymorphisms in HMGB1 may be a novel biomarker for female lung adenocarcinoma risk. PMID: 29617336
  35. Glycyrrhizin reduced the activity of JAK/STAT signaling pathway, which is the upstream regulator of HMGB1. PMID: 29568761
  36. HMGB1 may be a useful prognostic biomarker in malignant pleural mesothelioma when detected by immunohistochemistry, but cannot be considered a diagnostic biomarker in histological samples of mesothelioma PMID: 29356044
  37. the present study indicates that PBX 3'UTR may induce inflammatory responses and sepsis via acting as a competing endogenous RNA for HMGB1. PMID: 29484406
  38. overexpression of HMGB1 potentially promoted epileptogenesis. CLinduced activation of glial cells may act via upregulation of HMGB1 and TLR4/RAGE receptors, and the downstream transcription factor NFkappaB. PMID: 29393419
  39. Results show that HMGB1 is highly expressed in osteosarcoma tissues. Its expression is negatively regulated by miR-505 inhibiting proliferation, migration and invasion in osteosarcoma cells. PMID: 29251324
  40. This review demonstrated the increased expression of HMGB-1 in thrombosis-related diseases, including coronary artery disease, stroke, peripheral arterial disease, disseminated intravascular coagulation, and venous thrombosis. PMID: 29940562
  41. this study shows elevated serum level of HMGB1 in patients with the antiphospholipid syndrome PMID: 29410969
  42. Chronic periodontitis nonsmokers had elevated levels of HMGB1 in gingival crevicular fluid. The levels of HMGB1 were correlated with severity of periodontitis. Chronic periodontitis smokers exhibited lower levels of HMGB1 than chronic periodontitis nonsmokers. PMID: 28209360
  43. Serum HMGB1 levels in patients with inflamed appendix were significantly higher than in patients with normal appendix, and this could be a useful biomarker in improving the diagnostic accuracy of appendicitis. PMID: 27922766
  44. HMGB1 silencing promoted the susceptibility of retinoblastoma cells to chemotherapeutic drugs through downregulating NF-kappaB. PMID: 29328447
  45. HMGB1 possesses beneficial actions, such as endothelial activation, enhancement of neurite outgrowth, and neuronal survival in ischemic stroke. [review] PMID: 29054968
  46. HMGB1 mediates fibroblast activity via RAGE-MAPK and NF-kappaB signaling in keloid scar formation. PMID: 29283384
  47. Serum HMGB1 may be a potential marker to monitor the surgical course in patients undergoing surgery for colorectal cancer. PMID: 27834305
  48. the promotive effects of HuR overexpression on the inflammatory response were attenuated when HUVECs were cotreated with HMGB1 short hairpin RNA. Therefore, the present results indicated that the ectopic expression of HuR may induce inflammatory responses and thus sepsis by activating the HMGB1 signaling pathway. PMID: 29115544
  49. This is the first report to examine the risk factors associated with HMGB1 SNPs in the development of Rheumatoid arthritis disease in the Chinese Han population. PMID: 29200952
  50. HMGB1/IL-1beta complexes released after burn injuries can modulate immune responses PMID: 29601597

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Subcellular Location Nucleus, Chromosome, Cytoplasm, Secreted, Cell membrane, Peripheral membrane protein, Extracellular side, Endosome, Endoplasmic reticulum-Golgi intermediate compartment
Protein Families HMGB family
Tissue Specificity Ubiquituous. Expressed in platelets (PubMed:11154118).
Database Links

HGNC: 4983

OMIM: 163905

KEGG: hsa:3146

STRING: 9606.ENSP00000343040

UniGene: Hs.434102

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