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Human HMGB-1(High Mobility Group Protein B1) ELISA Kit

  • Cat.No.:E-EL-H1554

  • Reactivity: Human

To Purchase E-EL-H1554

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $609
Qty:

Product Details

Properties

Assay type Sandwich-ELISA
Format 96T/48T
Assay time 3.5h
Detection range 31.25-2000 pg/mL
Sensitivity 18.75 pg/mL
Sample type &Sample volume serum, plasma and other biological fluids; 100μL
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 < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of Human HMGB-1 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human HMGB-1. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human HMGB-1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human HMGB-1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human HMGB-1. You can calculate the concentration of Human HMGB-1 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
ItemSpecificationsStorage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips
-20℃, 6 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 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
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8°C
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human HMGB-1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human HMGB-1 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 99.32 202.75 693.26 108.96 208.03 746.59
Standard deviation 5.84 8.62 35.56 5.86 9.76 26.06
CV (%) 5.88 4.25 5.13 5.38 4.69 3.49

Recovery

The recovery of Human HMGB-1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum(n=8) 94-108 100
EDTA plasma (n=8) 94-107 100
Cell culture media (n=8) 93-105 99

Linearity

Samples were spiked with high concentrations of Human HMGB-1 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database LinksSwissProt:   P09429
SynonymsHMGB1, HMG1, HMG3, SBP-1, Amphoterin
Research AreaCancer, Epigenetics and Nuclear Signaling

Assay Procedures

elisa assay procedure 1

1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C

elisa assay procedure 2

2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C

elisa assay procedure 3

3. Aspirate and wash the plate for 3 times

elisa assay procedure 4

4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 5

5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 6

6. Add 50μL Stop Solution

elisa assay procedure 7

7. Read the plate at 450nm immediately. Calculation of the results

Citations

  1. ASN Neuro (2022) IF: 5.2
    Translocation of High Mobility Group Box 1 From the Nucleus to the Cytoplasm in Depressed Patients With Epilepsy

    DOI: 10.1177/17590914221136662

    Sample: Plasma
  2. Nanomedicine (2019) IF: 4.717
    Doxorubicin conjugated with nanodiamonds and in free form commit glioblastoma cells to heterodromous fates

    DOI: 10.2217/nnm-2018-0330

    PMID: 30676239

    Sample: Cell Culture Medium
  3. Indian Journal of Hematology and Blood Transfusion (2022) IF: 0.915
    Effects of AGEs, sRAGE and HMGB1 on Clinical Outcomes in Multiple MyelomaIn the PDF, in Header of all pages, Journal title should be abbreviated as "Indian J Hematol Blood Transfus".

    DOI: 10.1007/s12288-022-01574-6

    Sample: Serum
  4. Genetic Testing and Molecular Biomarkers (2019) IF: 1.121
    Association of High Mobility Group Box Protein B1 Gene Polymorphisms with Pneumonia Susceptibility and Severity

    DOI: 10.1089/gtmb.2018.0174

    PMID: 30562142

    Sample: Plasma
  5. Experimental and Therapeutic Medicine (2017) IF: 1.261
    Blood purification treatment initiated at the time of sepsis diagnosis effectively attenuates serum HMGB1 upregulation and improves patient prognosis

    DOI: 10.3892/etm.2017.4854

    Sample: Serum,Urine
  6. EPILEPTIC DISORDERS (2020) IF: 1.276
    HMGB‐1, TLR4, IL‐1R1, TNF‐α, and IL‐1β: novel epilepsy markers?

    DOI: 10.1684/epd.2020.1155

    Sample: Serum
  7. Experimental and Therapeutic Medicine (2018) IF: 1.41
    Necrosis of osteosarcoma cells induces the production and release of high‑mobility group box 1 protein

    DOI: 10.3892/etm.2017.5415

    Sample: Cell Culture Supernatant
  8. BIOLOGICAL & PHARMACEUTICAL BULLETIN (2019) IF: 1.54
    Triptolide suppresses growth of breast cancer by targeting HMGB1 in vitro and in vivo

    DOI: 10.1248/bpb.b18-00818

    PMID: 30956264

    Sample: Cell Culture Supernatant
  9. BIOCHEMICAL GENETICS (2022) IF: 2.22
    Isoquercitrin Induces Endoplasmic Reticulum Stress and Immunogenic Cell Death in Gastric Cancer Cells

    DOI: 10.1007/s10528-022-10309-1

    Sample: Gastric Cancer (Gc) Cells
  10. INTERNATIONAL UROLOGY AND NEPHROLOGY (2021) IF: 2.37
    High mobility group box 1 and homocysteine as preprocedural predictors for contrast-induced acute kidney injury after percutaneous coronary artery intervention

    DOI: 10.1007/s11255-021-03050-y

    PMID: 34727314

    Sample: Serum
  11. PeerJ (2020) IF: 2.379
    Glycyrrhizin suppresses epithelial-mesenchymal transition by inhibiting high-mobility group box1 via the TGF-β1/Smad2/3 pathway in lung epithelial cells

    DOI: 10.7717/peerj.8514

    Sample: Cell Culture Supernatant
  12. CLINICAL BIOCHEMISTRY (2019) IF: 2.43
    HMGB1 and SEPP1 as predictors of hepatocellular carcinoma in patients with viral C hepatitis: Effect of DAAs

    DOI: 10.1016/j.clinbiochem.2019.05.017

    PMID: 31158358

    Sample: Serum
  13. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS (2017) IF: 2.466
    Relationships of BRAF mutation and HMGB1 to papillary thyroid carcinoma

    DOI: 10.1016/j.bbrc.2017.03.117

    Sample: Serum
  14. Thoracic Cancer (2018) IF: 2.569
    Recombined humanized endostatin‐induced suppression of HMGB1 expression inhibits proliferation of NSCLC cancer cells

    DOI: 10.1111/1759-7714.12905

    Sample: Cell Culture Supernatant
  15. DNA AND CELL BIOLOGY (2016) IF: 2.574
    HMGB1 Enhances Drug Resistance and Promotes In Vivo Tumor Growth of Lung Cancer Cells

    DOI: 10.1089/dna.2016.3360

    Sample: Serum,Cell Culture Supernatant
  16. METABOLIC BRAIN DISEASE (2020) IF: 2.726
    Chronic fatigue syndrome and fibromyalgia-like symptoms are an integral component of the phenome of schizophrenia: neuro-immune and opioid system correlates

    DOI: 10.1007/s11011-020-00619-x

    PMID: 32965599

    Sample: Serum
  17. BMB Reports (2018) IF: 3.085
    29-kDa FN-f inhibited autophagy through modulating localization of HMGB1 in human articular chondrocytes

    DOI: 10.5483/BMBRep.2018.51.10.058

    Sample: Chondrocytes
  18. BRAIN RESEARCH BULLETIN (2019) IF: 3.103
    Role of toll-like receptor 4 in the regulation of the cell death pathway and neuroinflammation

    DOI: 10.1016/j.brainresbull.2019.03.015

    PMID: 30940475

    Sample: Cell Culture Medium
  19. JOURNAL OF NEUROIMMUNOLOGY (2020) IF: 3.125
    The association between plasma HMGB1 and sRAGE and clinical outcome in intracerebral hemorrhage

    DOI: 10.1016/j.jneuroim.2020.577266

    Sample: Plasma
  20. Frontiers in Physiology (2019) IF: 3.201
    Evidence for SIRT1 Mediated HMGB1 Release From Kidney Cells in the Early Stages of Hemorrhagic Shock

    DOI: 10.3389/fphys.2019.00854

    Sample: Serum
  21. MEDIATORS OF INFLAMMATION (2017) IF: 3.232
    Short-Term Postoperative Cognitive Dysfunction and Inflammatory Response in Patients Undergoing Cytoreductive Surgery and Hyperthermic Intraperitoneal Chemotherapy: A Pilot Study

    DOI: 10.1155/2017/3605350

    Sample: Serum
  22. SHOCK (2021) IF: 3.454
    The Value of Extracellular Cold-Inducible RNA-Binding Protein (eCIRP) in Predicting the Severity and Prognosis of Patients After Cardiac Arrest: A Preliminary Observational Study

    DOI: 10.1097/SHK.0000000000001702

    PMID: 34276038

    Sample: Serum
  23. TOXICOLOGY (2015) IF: 3.621
    Ethanol directly induced HMGB1 release through NOX2/NLRP1 inflammasome in neuronal cells

    DOI: 10.1016/j.tox.2015.06.006

    Sample: Cell Culture Supernatant
  24. Life-Basel (2022) IF: 3.817
    Response of Circulating Inflammatory Markers to Intermittent Hypoxia-Hyperoxia Training in Healthy Elderly People and Patients with Mild Cognitive Impairment

    DOI: 10.3390/life12030432

    Sample: Plasma
  25. American Journal of Translational Research (2019) IF: 4.06
    HMGB1-induced endothelial cell pyroptosis is involved in systemic inflammatory response syndrome following radiofrequency ablation of hepatic hemangiomas

    PMID: 31934300

    Sample: Serum,Huvecs
  26. Drug Design Development and Therapy (2021) IF: 4.162
    Prunella vulgaris L. Attenuates Experimental Autoimmune Thyroiditis by Inhibiting HMGB1/TLR9 Signaling

    DOI: 10.2147/DDDT.S325814

    PMID: 34764638

    Sample: Nthy-Ori 3–1 Cells
  27. Cells (2020) IF: 4.366
    Role of HMGB1/TLR4 Axis in Ischemia/Reperfusion-Impaired Extracellular Glutamate Clearance in Primary Astrocytes

    DOI: 10.3390/cells9122585

    PMID: 33287126

    Sample: Cell Culture Supernatant
  28. BIOCHEMICAL PHARMACOLOGY (2017) IF: 4.581
    Synthetic anti-endotoxin peptides inhibit cytoplasmic LPS-mediated responses

    DOI: 10.1016/j.bcp.2017.05.015

    Sample: Cell Culture Supernatant
  29. ADDICTION BIOLOGY (2017) IF: 4.603
    Young alcohol binge drinkers have elevated blood endotoxin, peripheral inflammation and low cortisol levels: neuropsychological correlations in women

    DOI: 10.1111/adb.12543

    Sample: Plasma
  30. JOURNAL OF CELLULAR AND MOLECULAR MEDICINE (2019) IF: 4.658
    Cigarette smoke‐induced HMGB1 translocation and release contribute to migration and NF‐κB activation through inducing autophagy in lung macrophages

    DOI: 10.1111/jcmm.14789

    Sample: Serum,Cell Culture Supernatant
  31. Nanomedicine (2017) IF: 4.727
    Monocyte-mediated chemotherapy drug delivery in glioblastoma

    DOI: 10.2217/nnm-2017-0266

    Sample: Supernatant
  32. JOURNAL OF AFFECTIVE DISORDERS (2021) IF: 4.839
    In schizophrenia, immune-inflammatory pathways are strongly associated with depressive and anxiety symptoms, which are part of a latent trait which comprises neurocognitive impairments and schizophrenia symptoms.

    DOI: 10.1016/j.jad.2021.03.062

    Sample: Serum
  33. Cell Communication and Signaling (2022) IF: 5.712
    The ROCK-ezrin signaling pathway mediates LPS-induced cytokine production in pulmonary alveolar epithelial cells

    DOI: 10.1186/s12964-022-00879-3

    PMID: 35551614

    Sample: A549 Cell
  34. Cancer Cell International (2021) IF: 5.722
    Extracellular HMGB1 interacts with RAGE and promotes chemoresistance in acute leukemia cells

    DOI: 10.1186/s12935-021-02387-9

    PMID: 34933679

    Sample: Jurkat Cell,Hl-60 Cell
  35. Frontiers in Oncology (2021) IF: 6.244
    CD68+ Macrophage Infiltration Associates With Poor Outcome of HPV Negative Oral Squamous Carcinoma Patients Receiving Radiation: Poly(I:C) Enhances Radiosensitivity of CAL-27 Cells but Promotes Macrophage Recruitment Through HMGB1

    DOI: 10.3389/fonc.2021.740622

    Sample: Cal-27 Cell
  36. JOURNAL OF HEADACHE AND PAIN (2021) IF: 7.277
    HMGB1, NLRP3, IL-6 and ACE2 levels are elevated in COVID-19 with headache: a window to the infection-related headache mechanism

    DOI: 10.1186/s10194-021-01306-7

    PMID: 34384355

    Sample: Serum
  37. Frontiers in Immunology (2018) IF: 7.561
    Profiling of Canonical and Non-Traditional Cytokine Levels in Interferon-β-Treated Relapsing–Remitting-Multiple Sclerosis Patients

    DOI: 10.3389/fimmu.2018.01240

    Sample: Serum
  38. JOURNAL OF CONTROLLED RELEASE (2017) IF: 7.786
    Harnessing the cross-talk between tumor cells and tumor-associated macrophages with a nano-drug for modulation of glioblastoma immune microenvironment

    DOI: 10.1016/j.jconrel.2017.10.024

    Sample: Cell Culture Supernatant
  39. LIVER INTERNATIONAL (2022) IF: 8.754
    Human poliovirus receptor contributes to biliary atresia pathogenesis by exacerbating natural-killer-cell-mediated bile duct injury

    DOI: 10.1111/liv.15457

    PMID: 36251580

    Sample: Serum
  40. BIOMATERIALS (2018) IF: 8.806
    Dendritic cell-mediated delivery of doxorubicin-polyglycerol-nanodiamond composites elicits enhanced anti-cancer immune response in glioblastoma

    DOI: 10.1016/j.biomaterials.2018.07.035

    Sample: Cell Culture Medium
  41. SCHIZOPHRENIA BULLETIN (2021) IF: 9.306
    High Mobility Group Protein 1 and Dickkopf-Related Protein 1 in Schizophrenia and Treatment-Resistant Schizophrenia: Associations With Interleukin-6, Symptom Domains, and Neurocognitive Impairments

    DOI: 10.1093/schbul/sbaa136

    Sample: Serum
  42. Journal of Neuroinflammation (2022) IF: 9.587
    Transient post-operative overexpression of CXCR2 on monocytes of traumatic brain injury patients drives monocyte chemotaxis toward cerebrospinal fluid and enhances monocyte-mediated immunogenic cell death of neurons in vitro

    DOI: 10.1186/s12974-022-02535-6

    PMID: 35768823

    Sample: Cerebrospinal Fluid (Csf),Peripheral Monocytes
  43. JOURNAL OF NANOBIOTECHNOLOGY (2021) IF: 10.435
    Synergy of nanodiamond–doxorubicin conjugates and PD-L1 blockade effectively turns tumor-associated macrophages against tumor cells

    DOI: 10.1186/s12951-021-01017-w

    Sample: A549 Cell,Lewis Cell
  44. Acta Pharmaceutica Sinica B (2022) IF: 14.903
    Reversing the PAI-1-induced fibrotic immune exclusion of solid tumor by multivalent CXCR4 antagonistic nano-permeator

    DOI: 10.1016/j.apsb.2022.12.012

    Sample: 4T1 Cells
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