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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

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

Related Products

Elabscience® provides ancillary reagents required for ELISA kit developments.11 reagents in total, including reagents for coating, blocking, washing, improving the stability and reducing matrix interference etc.
Cat.No. Product Name Specifications
E-ELIR-001 ELISA Plate Coating Buffer(1×) 100mL, 1000mL
E-ELIR-002 ELISA Plate Coating Buffer(5×) 100mL, 1000mL
E-ELIR-003 ELISA Plate Blocking Buffer 100mL, 1000mL
E-ELIR-004 Wash Buffer for Sandwich-ELISA(25×) 100mL, 1000mL
E-ELIR-005 Wash Buffer for Competitive-ELISA(25×) 100mL, 1000mL
E-ELIR-006 Stop Solution 100mL, 1000mL
E-ELIR-007 HRP-conjugate Stabilizer 10mL, 100mL, 500mL
E-ELIR-008 HRP-conjugate Diluent 100mL, 1000mL
E-ELIR-009 Biotinylated Antibody Stabilizer 10mL, 100mL, 500mL
E-ELIR-0010 Biotinylated Antibody Diluent 10mL, 100mL, 500mL
E-ELIR-0011 Sample Diluent 10mL, 100mL, 500mL

Citations

From now on, if you have published a paper by using any of our products since 1/1/2019, fill out the “Elabscience Publication Reward Application Form”carefully and send it to orders@elabscience.com, we will get back to you with the reward after we confirm it ASAP!

View more details about our Publication Reward >>

  1. Biomaterials (2018) IF: 8.805999999
    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

    PMID: 30071380

    Sample: Cell culture medium
  2. 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

    PMID: 29051064

    Sample: Cell culture supernatant
  3. Nanomedicine (2019) IF: 5.005
    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
  4. Frontiers in Immunology (2018) IF: 5.511
    Profiling of Canonical and Non-Traditional Cytokine Levels in Interferon-β-Treated Relapsing–Remitting-Multiple Sclerosis Patients.

    DOI: 10.3389/fimmu.2018.01240

    PMID: 29915590

    Sample: Serum
  5. 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

    PMID: 28840951

    Sample: Plasma
  6. Biochemical Pharmacology (2017) IF: 4.581
    Synthetic Anti-Endotoxin Peptides Inhibit Cytoplasmic LPS-Mediated Responses.

    DOI: 10.1016/j.bcp.2017.05.015

    PMID: 28539262

    Sample: Cell culture supernatant
  7. 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

    PMID: 99999999

    Sample: Plasma
  8. Brain Research Bulletin (2019) IF: 3.44
    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
  9. 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

    PMID: 29249869

    Sample: Serum
  10. Toxicology (2015) IF: 3.582
    Ethanol Directly Induced HMGB1 Release Through NOX2/NLRP1 Inflammasome In Neuronal Cells.

    DOI: 10.1016/j.tox.2015.06.006

    PMID: 26079697

    Sample: Cell culture supernatant
  11. Clinical biochemistry (2019) IF: 2.573
    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
  12. PeerJ (2020) IF: 2.379
    Glycyrrhizin suppresses epithelial-mesenchymal transition by inhibiting high-mobility group box1 via the TGF- β 1/Smad2/3 pathway in lung epithelial cells

    DOI: 10.7717/peerj.8514

    PMID: 32117622

    Sample: Cell culture supernatant
  13. Thoracic cancer (2019) IF: 2.569
    Recombined humanized endostatin-induced suppression of HMGB1 expression inhibits proliferation of NSCLC cancer cells

    DOI: 10.1111/1759-7714.12905

    PMID: 30485686

    Sample: Cell culture supernatant
  14. Biochemical And Biophysical Research Communications (2017) IF: 2.466
    Relationships Of BRAF Mutation And HMGB1To Papillary Thyroid Carcinoma.

    DOI: 10.1016/j.bbrc.2017.03.117

    PMID: 28342873

    Sample: Serum
  15. DNA and Cell Biology (2016) IF: 2.236
    Hmgb1 Enhances Drug Resistance And Promotes In Vivo Tumor Growth Of Lung Cancer Cells.

    DOI: 10.1089/dna.2016.3360

    PMID: 27383136

    Sample: Serum,Cell culture supernatant
  16. Genetic testing and molecular biomarkers (2018) IF: 1.181
    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
  17. Biological and Pharmaceutical Bulletin (2019) IF: 1.694
    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
  18. 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

    PMID: 28912856

    Sample: Serum,Urine
  19. Experimental and Therapeutic Medicine (2018) IF: 1.261
    Necrosis Of Osteosarcoma Cells Induces The Production And Release Of HighMobility Group Box 1 Protein.

    DOI: 10.3892/etm.2017.5415

    PMID: 29387198

    Sample: Cell culture supernatant
  20. Journal of Cellular and Molecular Medicine (2020)
    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

    PMID: 31769590

    Sample: Serum,Cell culture supernatant
  21. Pediatric Allergy (2018) IF: 0.533
    High-Mobility Group Protein Box 1 is Upregulated in Children with Henoch-Schonlein Purpura.

    DOI: 10.1089/ped.2018.0872

    Sample: Serum
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