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

    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience

      Catalog number:E-EL-H1554

      Synonyms:HMGB1, HMG1, HMG3, SBP-1, Amphoterin

      Size:
      • 96T
      • 24T
      Qty:
      - +
      Price: $609

      Reactivity: Human

      Detection Range: 31.25~2000 pg/mL

      Sensitivity: 18.75 pg/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request

      Intended use

      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.

      Assay type Sandwich
      Format 96T
      Assay time 3.5h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 31.25—2000 pg/mL
      Sensitivity 18.75 pg/mL
      Sample Volume Required Per Well 100μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Human HMGB-1 in samples. No significant cross-reactivity or interference between Human HMGB-1 and analogues was observed.

      Typical data

      As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

      Concentration
      (pg/mL)
      O.D Average Corrected
      2000 2.443
      2.447
      2.445 2.374
      1000 1.718
      1.764
      1.741 1.67
      500 0.988
      0.984
      0.986 0.915
      250 0.466
      0.498
      0.482 0.411
      125 0.265
      0.247
      0.256 0.185
      62.5 0.187
      0.161
      0.174 0.103
      31.25 0.118
      0.13
      0.124 0.053
      0 0.069
      0.073
      0.071 --

      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
      C V (%) 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=5) 94-108 100
      EDTA plasma (n=5) 94-107 100
      Cell culture media (n=5) 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.

      Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
      1:2 Range (%) 98-111 92-105 85-100
      Average (%) 105 98 92
      1:4 Range (%) 90-103 84-100 88-102
      Average (%) 98 91 94
      1:8 Range (%) 87-99 83-93 85-96
      Average (%) 94 88 90
      1:16 Range (%) 88-103 85-96 86-99
      Average (%) 94 90 91

      Kit components & Storage

      An unopened kit can be stored at 2-8°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

      Item Specifications Storage
      Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20°C, 6 months
      Reference Standard 2 vials
      Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
      Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20°C(shading 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°C(shading light)
      Stop Solution 1 vial, 10 mL 2-8°C
      Plate Sealer 5 pieces
      Product Description 1 copy
      Certificate of Analysis 1 copy

      Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
      The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

      Other supplies required

      • Microplate reader with 450 nm wavelength filter
      • High-precision transfer pipette, EP tubes and disposable pipette tips
      • Incubator capable of maintaining 37°C
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      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. Ethanol Directly Induced HMGB1 Release Through NOX2/NLRP1 Inflammasome In Neuronal Cells.
        IF: 3.582
        Journal: Toxicology(2015)
        DOI: 10.1016/j.tox.2015.06.006
        PMID: 26079697
        Products: E-EL-H1554 E-EL-R0505 
        Sample type: Cell culture supernatant
        Reactivity: Human,Rat
      2. Hmgb1 Enhances Drug Resistance And Promotes In Vivo Tumor Growth Of Lung Cancer Cells.
        IF: 2.236
        Journal: DNA and Cell Biology(2016)
        DOI: 10.1089/dna.2016.3360
        PMID: 27383136
        Products: E-EL-H1554 
        Sample type: Serum,Cell culture supernatant
        Reactivity: Human
      3. Relationships Of BRAF Mutation And HMGB1To Papillary Thyroid Carcinoma.
        IF: 2.466
        Journal: Biochemical And Biophysical Research Communications(2017)
        DOI: 10.1016/j.bbrc.2017.03.117
        PMID: 28342873
        Products: E-EL-H1554 
        Sample type: Serum
        Reactivity: Human
      4. Synthetic Anti-Endotoxin Peptides Inhibit Cytoplasmic LPS-Mediated Responses.
        IF: 4.581
        Journal: Biochemical Pharmacology(2017)
        DOI: 10.1016/j.bcp.2017.05.015
        PMID: 28539262
        Products: E-EL-H1554 
        Sample type: Cell culture supernatant
        Reactivity: Human
      5. Necrosis Of Osteosarcoma Cells Induces The Production And Release Of HighMobility Group Box 1 Protein.
        IF: 1.261
        Journal: Experimental and Therapeutic Medicine(2018)
        DOI: 10.3892/etm.2017.5415
        PMID: 29387198
        Products: E-EL-H1554 
        Sample type: Cell culture supernatant
        Reactivity: Human
      6. Young Alcohol Binge Drinkers Have Elevated Blood Endotoxin, Peripheral Inflammation And Low Cortisol Levels: Neuropsychological Correlations In Women.
        IF: 4.603
        Journal: Addiction Biology(2017)
        DOI: 10.1111/adb.12543
        PMID: 28840951
        Products: E-EL-H1554 E-EL-H6005 
        Sample type: Plasma
        Reactivity: Human
      7. Short-Term Postoperative Cognitive Dysfunction And Inflammatory Response In Patients Undergoing Cytoreductive Surgery And Hyperthermic Intraperitoneal Chemotherapy: A Pilot Study.
        IF: 3.232
        Journal: Mediators Of Inflammation(2017)
        DOI: 10.1155/2017/3605350
        PMID: 29249869
        Products: E-EL-H0149 E-EL-H0109 E-EL-H1554 E-EL-H2183 E-EL-H1297 
        Sample type: Serum
        Reactivity: Human
      8. Harnessing The Cross-Talk Between Tumor Cells And Tumor-Associated Macrophages With A Nano-Drug For Modulation Of Glioblastoma Immune Microenvironment.
        IF: 7.786
        Journal: Journal of Controlled Release(2017)
        DOI: 10.1016/j.jconrel.2017.10.024
        PMID: 29051064
        Products: E-EL-H1554 
        Sample type: Cell culture supernatant
        Reactivity: Human
      9. Blood Purification Treatment Initiated At The Time Of Sepsis Diagnosis Effectively Attenuates Serum HMGB1 Upregulation And Improves Patient Prognosis.
        IF: 1.261
        Journal: Experimental and Therapeutic Medicine(2017)
        DOI: 10.3892/etm.2017.4854
        PMID: 28912856
        Products: E-EL-H1554 
        Sample type: Serum,Urine
        Reactivity: Human
      10. Dendritic cell-mediated delivery of doxorubicin-polyglycerol-nanodiamond composites elicits enhanced anti-cancer immune response in glioblastoma.
        IF: 8.805999999
        Journal: Biomaterials(2018)
        DOI: 10.1016/j.biomaterials.2018.07.035
        PMID: 30071380
        Products: E-EL-H1554 
        Sample type: Cell culture medium
        Reactivity: Human
      11. High-Mobility Group Protein Box 1 is Upregulated in Children with Henoch-Schonlein Purpura.
        IF: 0.533
        Journal: Pediatric Allergy(2018)
        DOI: 10.1089/ped.2018.0872
        Products: E-EL-H1554 E-EL-H0102 E-EL-H6008 E-EL-H0109 
        Sample type: Serum
        Reactivity: Human
      12. Profiling of Canonical and Non-Traditional Cytokine Levels in Interferon-β-Treated Relapsing–Remitting-Multiple Sclerosis Patients.
        IF: 5.511
        Journal: Frontiers in Immunology(2018)
        DOI: 10.3389/fimmu.2018.01240
        PMID: 29915590
        Products: E-EL-H1554 
        Sample type: Serum
        Reactivity: Human
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