Human HMGB1 ELISA Kit

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Antigen
High-Mobility Group Box 1 (HMGB1) ELISA Kits
  • hmgb1
  • amphoterin
  • hmg-1
  • hmg1
  • hmg3
  • sbp-1
  • HMGB1
  • HMG1
  • HMG3
  • SBP-1
  • DEF
  • HMG-1
  • Hmg1
  • p30
  • Ac2-008
  • ik:tdsubc_1a5
  • wu:fb23c02
  • xx:tdsubc_1a5
  • zgc:56110
  • zgc:77104
  • Hmgb1
  • high-mobility group box 1
  • high mobility group box 1
  • high mobility group protein B1-like
  • high-mobility group box 1a
  • HMG-box transcription factor 1
  • hmgb1
  • HMGB1
  • Hmgb1
  • LOC100392312
  • LOC100754755
  • LOC100359149
  • hmgb1a
  • Hbp1
Reactivity
Human
Alternatives
Kits with alternative reactivity to:
16
13
13
8
7
5
3
2
2
1
1
1
1
1
Method Type
Sandwich ELISA
Detection Range
62.5-4000 pg/mL
Minimum Detection Limit
62.5 pg/mL
Application
ELISA
Options
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Catalog No. ABIN414391
$ 530.53
Plus shipping costs $45.00
Relevance Score ABIN Method Type Sample Type Detection Range Detection Minimum Supplier References Details
12.499232 ABIN1570312 Sandwich ELISA Biological Fluids, Plasma, Serum 62.5-4000 pg/mL 62.5 pg/mL Log in to see
12.499232 ABIN368346 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate 78-5000 pg/mL 78 pg/mL Log in to see
12.499232 ABIN1081530 Sandwich ELISA Serum, Plasma, Biological Fluids 13.7-10.000 pg/mL 13.7 pg/mL Log in to see
12.499232 ABIN5564695 Sandwich ELISA Biological Fluids, Plasma, Serum 62.5-4000 pg/mL 62.5 pg/mL Log in to see
1 ABIN455352 Sandwich ELISA Cell Culture Supernatant, Serum, Plasma, Biological Fluids 0.156-10 ng/mL 0.156 ng/mL Log in to see 2
1 ABIN511375 Competition ELISA Serum, Cell Culture Supernatant, Tissue Homogenate, Plasma, Body Fluids 2.5-50 ng/mL 2.5 ng/mL Log in to see 1
1 ABIN5593802 Sandwich ELISA Biological Fluids, Plasma, Serum, Tissue Homogenate 0.156-10 ng/mL 0.156 ng/mL Log in to see
1 ABIN5523371 Sandwich ELISA Plasma, Serum, Biological Fluids, Tissue Homogenate 31.25-2000 pg/mL 31.25 pg/mL Log in to see
1 ABIN5519093 Sandwich ELISA Plasma, Serum, Biological Fluids 31.25-2000 pg/mL 31.25 pg/mL Log in to see
1 ABIN2087229 Sandwich ELISA 2.5-50 ng/mL 2.5 ng/mL Log in to see
1 ABIN2534491 Sandwich ELISA Serum, Plasma, Biological Fluids 62.5-4000 pg/mL 62.5 pg/mL Log in to see
1 ABIN4969872 Sandwich ELISA Cell Culture Supernatant, Plasma, Tissue Homogenate, Serum, Cell Lysate, Biological Fluids 62.5-4000 pg/mL 62.5 pg/mL Log in to see
1 ABIN2944225 13.72-10000 pg/mL 13.72 pg/mL Log in to see
1 ABIN2948301 31.25-2000 pg/mL 31.25 pg/mL Log in to see
1 ABIN5072222 Sandwich ELISA Plasma, Serum, Tissue Homogenate, Biological Fluids Log in to see
1 ABIN5074386 Log in to see

General

Antigen High-Mobility Group Box 1 (HMGB1) ELISA Kits
Reactivity Human
Kits with alternative reactivity to:
(16), (13), (13), (8), (7), (5), (3), (2), (2), (1), (1), (1), (1), (1)
Method Type Sandwich ELISA
Detection Range 62.5-4000 pg/mL
Minimum Detection Limit 62.5 pg/mL
Application ELISA
Pubmed 20 references available
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Product Details HMGB1 ELISA Kit

Target details Application Details Handling ProductDetails: References for HMGB1 Kit (ABIN414391) Images
Purpose The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of HMG1 in Serum,Plasma,Biological Fluids
Sample Type Serum, Plasma, Biological Fluids
Detection Method Colorimetric
Analytical Method Quantitative
Specificity

This assay has high sensitivity and excellent specificity for detection of High Mobility Group Protein 1 (HMG1).
No significant cross-reactivity or interference between High Mobility Group Protein 1 (HMG1) and analogues was observed.

Cross-Reactivity (Details) No significant cross-reactivity or interference between High Mobility Group Protein 1 (HMG1) and analogues was observed.
Sensitivity 28.3 pg/mL
Components
  • Pre-coated, ready to use 96-well strip plate
  • Plate sealer for 96 wells
  • Standard Diluent
  • Assay Diluent A
  • Assay Diluent B
  • Stop Solution
  • Standard
  • Detection Reagent A
  • Detection Reagent B
  • TMB Substrate
  • Wash Buffer (30 × concentrate)
  • Instruction manual
Material not included
  • Microplate reader with 450 nm filter.
  • Precision single or multi-channel pipettes and disposable tips.
  • Eppendorf Tubes for diluting samples.
  • Deionized or distilled water.
  • Absorbent paper for blotting the microtiter plate.
  • Container for Wash Solution

Target details

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Antigen
Alternative Name HMG1 (HMGB1 ELISA Kit Abstract)
UniProt P09429
Pathways p53 Signaling, Regulation of Muscle Cell Differentiation, Skeletal Muscle Fiber Development, Positive Regulation of Endopeptidase Activity, Regulation of Carbohydrate Metabolic Process, Toll-Like Receptors Cascades, Smooth Muscle Cell Migration

Application Details

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Application Notes
  • Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
  • The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
  • Kits from different batches may be a little different in detection range, sensitivity and color developing time.
  • Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
  • Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
  • There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
  • Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
  • Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
  • Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
  • Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.
Comment

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume 100 μL
Assay Time 3 h
Plate Pre-coated
Protocol The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to High Mobility Group Protein 1 (HMG1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to High Mobility Group Protein 1 (HMG1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain High Mobility Group Protein 1 (HMG1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of High Mobility Group Protein 1 (HMG1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Reagent Preparation
  • Bring all kit components and samples to room temperature (18-25°C) before use.
  • Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 4,000pg/mL. Then prepare 7 tubes containing 0.5mL Standard Diluent and use the standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, and the last EP tubes with Standard Diluent is the blank as 0pg/mL.
  • Assay Diluent A and Assay Diluent B - Dilute 6mL of Assay Diluent A or B Concentrate(2×) with 6mL of deionized or distilled water to prepare 12mL of Assay Diluent A or B. (In fact, more than 6mL Assay Diluent A and Assay Diluent B are contained in the bottles. Therefore, in every test, please precisely pipette required amount of Diluent and make double dilution in a new container. The prepared working dilution cannot be frozen.)
  • Detection Reagent A and Detection Reagent B - Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute to the working concentration with working Assay Diluent A or B, respectively (1:100).
  • Wash Solution - Dilute 20mL of Wash Solution concentrate (30×) with 580mL of deionized or distilled water to prepare 600mL of Wash Solution (1×).
  • TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.
Note:
  • Making serial dilution in the wells directly is not permitted.
  • Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37°C directly.
  • Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10µL for once pipetting.
  • The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
  • If crystals have formed in the Wash Solution concentrate (30×), warm to room temperature and mix gently until the crystals are completely dissolved.
  • Contaminated water or container for reagent preparation will influence the detection result.
Sample Collection Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000 × g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

Biological Fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles.
Sample Preparation
  • We are only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.
  • Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments. Sample should be diluted by 0.01mol/L PBS(PH=7.0-7.2).
  • If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is necessary.
  • Tissue or cell extraction samples prepared by chemical lysis buffer may cause unexpected ELISA results due to the impacts from certain chemicals.
  • Due to the possibility of mismatching between antigen from other origin and antibody used in our kits (e.g.antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
  • Influenced by the factors including cell viability, cell number or sampling time, samples from cell culture supernatant may not be detected by the kit.
  • Fresh samples without long time storage is recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.
Assay Procedure
  1. Prepare all reagents, samples and standards,
  2. Add 100μL standard or sample to each well. Incubate 2 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.
Calculation of Results Average the duplicate readings for each standard, control, and samples and subtract the average zero standard optical density. Construct a standard curve by plotting the mean O.D. and concentration for each standard and draw a best fit curve through the points on the graph or create a standard curve on log-log graph paper with HMG1 concentration on the y-axis and absorbance on the x-axis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the known concentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is the dependent variable. However, the O.D. values of the standard curve may vary according to the conditions of assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), plotting log of the data to establish standard curve for each test is recommended. Typical standard curve below is provided for reference only.
Assay Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Mobility Group Protein 1 (HMG1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Mobility Group Protein 1 (HMG1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions For Research Use only

Handling

Product Details HMGB1 ELISA Kit Target details Application Details ProductDetails: References for HMGB1 Kit (ABIN414391) Images back to top
Precaution of Use The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
Handling Advice

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.

Storage 4 °C
Storage Comment
  • For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
  • For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
    Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
  • For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.
Expiry Date 6 months

ProductDetails: References for HMGB1 Kit (ABIN414391)

Product Details HMGB1 ELISA Kit Target details Application Details Handling Images back to top
Product cited in:

Chen, Fang, Li, Chen, Li, Gong, Fang: "Glycyrrhizin ameliorates experimental colitis through attenuating interleukin-17-producing T cell responses via regulating antigen-presenting cells." in: Immunologic research, Vol. 65, Issue 3, pp. 666-680, 2017

Yin, Feng, Zhao, Zhao, Yua, Xu, Che: "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." in: Biomedicine & pharmacotherapy, Vol. 88, pp. 449-458, 2017

Gmiat, Mieszkowski, Prusik, Prusik, Kortas, Kochanowicz, Radulska, Lipiński, Tomczyk, Jaworska, Antosiewicz, Ziemann: "Changes in pro-inflammatory markers and leucine concentrations in response to Nordic Walking training combined with vitamin D supplementation in elderly women." in: Biogerontology, Vol. 18, Issue 4, pp. 535-548, 2017

Ying, Jiang, He, Yu, Chen, Chen, Ru, Chen, Chen, Zhu, Li, Zhang, Guo, Yin, Zhang, Lou: "Serum HMGB1 as a Potential Biomarker for Patients with Asbestos-Related Diseases." in: Disease markers, Vol. 2017, pp. 5756102, 2017

Kargı, Demirpençe, Gündüz, Göktaş, Alikanoǧlu, Yıldırım: "Serum levels of HMGB1 have a diagnostic role in metastatic renal cell cancer." in: Cancer biomarkers : section A of Disease markers, Vol. 17, Issue 1, pp. 17-20, 2016

Ming, Gao, Zou, Chen, Sun, Xiao, Lai, Xiong, Xu, Tan, Wang, Chen, Gong, Xia, Zheng: "HMGB1 blockade differentially impacts pulmonary inflammation and defense responses in poly(I:C)/LPS-exposed heart transplant mice." in: Molecular immunology, Vol. 76, pp. 80-9, 2016

Wang, Sun, Li, Deng, Zeng, Tao, Wang, Guan, Zhao: "Urinary MCP-1、HMGB1 increased in calcium nephrolithiasis patients and the influence of hypercalciuria on the production of the two cytokines." in: Urolithiasis, Vol. 45, Issue 2, pp. 159-175, 2016

Wang, Wei, Liu, Zhou, Xiao, Dong, Li: "The Role of High Mobility Group Box 1 Protein in Interleukin-18-Induced Myofibroblastic Transition of Valvular Interstitial Cells." in: Cardiology, Vol. 135, Issue 3, pp. 168-178, 2016

Wang, Li, Yu, Guo, Yang, Zhang, Li, Li, Hu, Zheng, Song: "High Mobility Group Box 1 Mediates Interferon-γ-Induced Phenotypic Modulation of Vascular Smooth Muscle Cells." in: Journal of cellular biochemistry, Vol. 118, Issue 3, pp. 518-529, 2016

Wang, Miao, Wu, Huang, Sun, Zhu, Zheng, Sun, Dong: "Serum HMGB1 Serves as a Novel Laboratory Indicator Reflecting Disease Activity and Treatment Response in Ankylosing Spondylitis Patients." in: Journal of immunology research, Vol. 2016, pp. 6537248, 2016

Basso, Padoan, Laufer, Aneloni, Moz, Schroers, Pelloso, Saiz, Krapp, Fogar, Cornoldi, Zambon, Rossi, La Malfa, Marotti, Brefort, Weis, Katus, Plebani: "Relevance of pre-analytical blood management on the emerging cardiovascular protein biomarkers TWEAK and HMGB1 and on miRNA serum and plasma profiling." in: Clinical biochemistry, Vol. 50, Issue 4-5, pp. 186-193, 2016

Alizadeh, Ghazavi, Ganji, Mosayebi: "Diagnostic Value of High-Mobility Group Box 1 (HMGB1) Protein in Acute and Perforated Appendicitis." in: Journal of investigative surgery : the official journal of the Academy of Surgical Research, pp. 1-5, 2016

Kwon, Kim, Park, Choi, Lee, Lee, Heo, Chang, Koh: "Increased VEGF and decreased SDF-1? in patients with silent brain infarction are associated with better prognosis after first-ever acute lacunar stroke." in: Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association, Vol. 24, Issue 3, pp. 704-10, 2015

Qin, Mi, Li, Wang, Zhang, Wang, Zhang, Ma, Wu, Zhang: "Low shear stress induced HMGB1 translocation and release via PECAM-1/PARP-1 pathway to induce inflammation response." in: PLoS ONE, Vol. 10, Issue 3, pp. e0120586, 2015

Qu, Wang, Huang, Qiu, Xiang, Lu: "High mobility group box 1 gene polymorphism is associated with the risk of postoperative atrial fibrillation after coronary artery bypass surgery." in: Journal of cardiothoracic surgery, Vol. 10, pp. 88, 2015

Wang, Pei, Zhu, Zhou, Liu, Xiong, Liu, Lin, Xie: "Overexpression of HMGB1 A-box reduced lipopolysaccharide-induced intestinal inflammation via HMGB1/TLR4 signaling in vitro." in: World journal of gastroenterology, Vol. 21, Issue 25, pp. 7764-76, 2015

Xiao, Wu, Yin, Han, Ding, Qiao, Lu, Deng, Bo, Gong: "Wogonin Inhibits Tumor-derived Regulatory Molecules by Suppressing STAT3 Signaling to Promote Tumor Immunity." in: Journal of immunotherapy (Hagerstown, Md. : 1997), Vol. 38, Issue 5, pp. 167-84, 2015

Wang, Qu, Deng: "Plasma HMGB-1 Levels in Subjects with Obesity and Type 2 Diabetes: A Cross-Sectional Study in China." in: PLoS ONE, Vol. 10, Issue 8, pp. e0136564, 2015

Young, Huang, Wu, Lai, Hou, Peng, Liang, Lin, Chang, Tsai, Wu, Wu, Ko: "Hemojuvelin modulates iron stress during acute kidney injury: improved by furin inhibitor." in: Antioxidants & redox signaling, Vol. 20, Issue 8, pp. 1181-94, 2014

Iwamoto, Gao, Pulkkinen, Toljamo, Nieminen, Mazur: "Soluble receptor for advanced glycation end-products and progression of airway disease." in: BMC pulmonary medicine, Vol. 14, pp. 68, 2014

Images

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Supplier Images
ELISA image for High-Mobility Group Box 1 (HMGB1) ELISA Kit (ABIN414391) High-Mobility Group Box 1 (HMGB1) ELISA Kit