HMGB1 ELISA Kit

Catalog No. ABIN6574155

Product Details HMGB1 ELISA Kit

Target: HMGB1 (High Mobility Group Box 1 (HMGB1))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5 pg/mL - 4000 pg/mL

Minimum Detection Limit: 62.5 pg/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of HMG1 in human serum, plasma.

We offer validation data (WB) for the kit components. So you can be sure to order a reliable ELISA kit product composed of high quality reagents.

Sample Type: Plasma, Serum

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of High Mobility Group Protein 1 (HMGB1)

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, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Product Specific Information:

What can the HMGB1 ELISA Kit ABIN6574155 be used for? This human HMGB1 ELISA kit detects High Mobility Group Protein 1 (HMGB1, syn. HMG1) by sandwich ELISA with high sensitivity and reliability within 3 hours. No significant cross-reactivity or interference between HMGB1 and the analogues could be observed. The detection range of this high quality validated HMGB1 ELISA kit is between 62.5 and 4000 pg/ml. The sensitivity is 28.3 pg/mL. The required sample volume is 100µL. The ELISA Kit for the detection of HMGB1 contains the relevant components and instructions and is easy to use.

What validation data is available for this HMGB1 ELISA Kit? The ELISA Kit has been used in 50 publications to date, which are listed below or can be viewed at Pubmed. The HMGB1 ELISA Kit has been extensively validated. Currently, 4 product images are available showing western blots of the standard and capture antibody included in the kit, as well as an SDS page of the standard. Use our HMGB1 ELISA Kit to reliably, easily and quickly detect HMGB1.

What is the function of HMGB1? HMGB1 or HMG1 is a multifunctional redox-sensitive protein with various roles in different cellular compartments. In the nucleus, it is one of the major chromatin-associated non-histone proteins and functions as a DNA chaperone involved in replication, transcription, chromatin remodeling, V(D)J recombination, DNA repair and genome stability (PubMed:33147444).

Application Details

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:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 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.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 0.5 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 4,000pg/mL. Prepare 7 tubes containing 0.25 mL Standard Diluent and produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 4,000pg/mL, 2,000pg/mL, 1,000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0pg/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. 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 one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target 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

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.

Storage: 4 °C/-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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Expiry Date: 6 months

References for HMGB1 ELISA Kit (ABIN6574155)

Angioni, Sasset, Cioccarelli, Sánchez-Rodríguez, Bertoldi, Putaggio, Viola, Cattelan, Molon: "COVID-19 Vaccination Limits Systemic Danger Signals in SARS-CoV-2 Infected Patients." in: , Vol. 14, Issue 3, (2022) (PubMed).

Wang, Li, Park, Gonzalez, Scott, Lu: "Camptothesome elicits immunogenic cell death to boost colorectal cancer immune checkpoint blockade." in: Journal of controlled release : official journal of the Controlled Release Society, Vol. 349, pp. 929-939, (2022) (PubMed).

Wang, Cordova, Chalasani, Lu: "Camptothesome Potentiates PD-L1 Immune Checkpoint Blockade for Improved Metastatic Triple-Negative Breast Cancer Immunochemotherapy." in: Molecular pharmaceutics, Vol. 19, Issue 12, pp. 4665-4674, (2022) (PubMed).

Wu, Liu, Zhang: "LINC00240/miR-155 axis regulates function of trophoblasts and M2 macrophage polarization via modulating oxidative stress-induced pyroptosis in preeclampsia." in: Molecular medicine (Cambridge, Mass.), Vol. 28, Issue 1, pp. 119, (2022) (PubMed).

Brajer-Luftmann, Nowicka, Kaczmarek, Wyrzykiewicz, Yasar, Piorunek, Grabicki, Kostrzewska, Sikora, Batura-Gabryel: "Molecules of Damage-Associated Patterns in Bronchoalveolar Lavage Fluid and Serum in Chronic Obstructive Pulmonary Disease." in: Advances in experimental medicine and biology, Vol. 1113, pp. 27-35, (2019) (PubMed).

Lin, Biswas, Choolani, Fong, Bongso: "Induction of Immunogenic Cell Death in Lymphoma Cells by Wharton's Jelly Mesenchymal Stem Cell Conditioned Medium." in: Stem cell reviews, Vol. 13, Issue 6, pp. 801-816, (2018) (PubMed).

Wang, Peng, Huang, Liu, Kong, Dong, Dai, Zhou, Wang, Yang, Cheng, Gao, Qu, Wang, Zhu, Tian, Liu, Cao, Cui, Xu, Xu, Sun: "Blocking the Feedback Loop between Neuroendocrine Differentiation and Macrophages Improves the Therapeutic Effects of Enzalutamide (MDV3100) on Prostate Cancer." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 24, Issue 3, pp. 708-723, (2018) (PubMed).

Salaroglio, Panada, Moiso, Buondonno, Provero, Rubinstein, Kopecka, Riganti: "PERK induces resistance to cell death elicited by endoplasmic reticulum stress and chemotherapy." in: Molecular cancer, Vol. 16, Issue 1, pp. 91, (2018) (PubMed).

Oh, Park, Lee, Seo, Shin, Oh et al.: "Assessing the recovery from prerenal and renal acute kidney injury after treatment with single herbal medicine via activity of the biomarkers HMGB1, NGAL and KIM-1 in kidney proximal tubular cells ..." in: BMC complementary and alternative medicine, Vol. 17, Issue 1, pp. 544, (2018) (PubMed).

Riganti, Lingua, Salaroglio, Falcomatà, Righi, Morena, Picca, Oddo, Kopecka, Pradotto, Libener, Orecchia, Bironzo, Comunanza, Bussolino, Novello, Scagliotti, Di Nicolantonio, Taulli: "Bromodomain inhibition exerts its therapeutic potential in malignant pleural mesothelioma by promoting immunogenic cell death and changing the tumor immune-environment." in: Oncoimmunology, Vol. 7, Issue 3, pp. e1398874, (2018) (PubMed).

Li, Huang, Wang, Zhang: "Increased serum levels of high mobility group protein B1 and calprotectin in pre-eclampsia." in: International journal of gynaecology and obstetrics: the official organ of the International Federation of Gynaecology and Obstetrics, Vol. 142, Issue 1, pp. 37-41, (2018) (PubMed).

Rojas-Sepúlveda, Tittarelli, Gleisner, Ávalos, Pereda, Gallegos, González, López, Butte, Roa, Fluxá, Salazar-Onfray: "Tumor lysate-based vaccines: on the road to immunotherapy for gallbladder cancer." in: Cancer immunology, immunotherapy : CII, Vol. 67, Issue 12, pp. 1897-1910, (2018) (PubMed).

Keshk, Zineldeen, El-Heneedy, Ghali: "Thrombomodulin, alarmin signaling, and copeptin: cross-talk between obesity and acute ischemic stroke initiation and severity in Egyptians." in: Neurological sciences : official journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology, Vol. 39, Issue 6, pp. 1093-1104, (2018) (PubMed).

Yan, Zhu, Zhang, Li, Li, Wang, Leng: "HMGB1-RAGE signaling pathway in pPROM." in: Taiwanese journal of obstetrics & gynecology, Vol. 57, Issue 2, pp. 211-216, (2018) (PubMed).

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) (PubMed).

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) (PubMed).

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) (PubMed).

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) (PubMed).

Hou, Luan, Ren: "Oxidized low-density lipoprotein promotes osteoclast differentiation from CD68 positive mononuclear cells by regulating HMGB1 release." in: Biochemical and biophysical research communications, Vol. 495, Issue 1, pp. 1356-1362, (2017) (PubMed).

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) (PubMed).

Target Details for HMGB1

Target: HMGB1 (High Mobility Group Box 1 (HMGB1))

Alternative Name: High Mobility Group Protein 1 (HMGB1) (HMGB1 Products)

Synonyms: HMG1 ELISA Kit, HMG3 ELISA Kit, SBP-1 ELISA Kit, DEF ELISA Kit, HMG-1 ELISA Kit, Hmg1 ELISA Kit, amphoterin ELISA Kit, p30 ELISA Kit, hmgb1 ELISA Kit, ik:tdsubc_1a5 ELISA Kit, wu:fb23c02 ELISA Kit, xx:tdsubc_1a5 ELISA Kit, zgc:56110 ELISA Kit, zgc:77104 ELISA Kit, hmg-1 ELISA Kit, hmg3 ELISA Kit, sbp-1 ELISA Kit, hmg1 ELISA Kit, HMGB1 ELISA Kit, Ac2-008 ELISA Kit, high mobility group box 1 ELISA Kit, high-mobility group box 1 ELISA Kit, high mobility group box 1a ELISA Kit, high mobility group box 1 L homeolog ELISA Kit, high mobility group protein B1 ELISA Kit, HMGB1 ELISA Kit, Hmgb1 ELISA Kit, hmgb1 ELISA Kit, hmgb1a ELISA Kit, hmgb1.L ELISA Kit, LOC100359149 ELISA Kit

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, Inflammasome