HMGB1 ELISA Kit

Produktnummer ABIN6962899

Maus HMGB1 ELISA Kit, Colorimetric Assay für die Quantifizierung von Maus HMGB1 und es wurde in 5+ Publikationen zitiert.

Kurzübersicht für HMGB1 ELISA Kit (ABIN6962899)

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

Reaktivität: Maus

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 15.63 pg/mL - 1000 pg/mL

Applikation: ELISA

Proben: Plasma, Serum

Produktdetails HMGB1 ELISA Kit

Untere Nachweisgrenze: 15.63 pg/mL

Verwendungszweck: The kit is a sandwich enzyme immunoassay technique for the in vitro quantitative measurement in various sample types.

Analytische Methode: Quantitative

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

Sensitivität: 9.38 pg/mL

Bestandteile:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Reference Standard & Sample Diluent
• Biotinylated Detection Antibody (100 x concentrate)
• HRP Conjugate (100 x concentrate)
• Biotinylated Detection Antibody Diluent
• HRP Conjugate Diluent
• Substrate Reagent
• Stop Solution
• Wash Buffer (25 x concentrate)
• Instruction manual

Anwendungsinformationen

Probenmenge: 100 μL

Testdauer: 3.5 h

Plattentyp: Pre-coated

Protokoll:

1. Add 100 µL standard or sample to each well. Incubate for 90 min at 37 °C.
2. Remove the liquid. Add 100 µL Biotinylated Detection Antibody. Incubate for 1 hour at 37 °C.
3. Aspirate and wash 3 times.
4. Add 100 µL HRP Conjugate. Incubate for 30 min at 37 °C.
5. Aspirate and wash 5 times.
6. Add 90 µL Substrate Reagent. Incubate for 15 min at 37 °C.
7. Add 50 µL Stop Solution. Read at 450 nm immediately.
8. Calculation of results.

Aufbereitung der Reagenzien:

1. Equilibrate all reagents to room temperature (18-25 °C) before use. If the kit will not be used up in one assay, please only take out the necessary strips and reagents for present experiment, and store the remaining strips and reagents at required condition.
2. Standard Working Solution: Centrifuge the standard at 10,000x g for 1 min. Reconstitute with 1 mL of the Standard & Sample Diluent, allow to stand for 10 minutes, and gently invert several times until completely dissolved. Alternatively, vortex briefly at low speed, remove bubbles by low-speed centrifugation if necessary.This produces a 1000 pg/mL stock solution. Prepare serial dilutions according to experimental requirements. Recommended dilution gradient: 1000, 500, 250, 125, 62.5, 31.25, 15.63, 0 pg/mL Serial Dilution Procedure: Dispense 500 μL of Standard & Sample Diluent into 7 EP tubes, respectively. Transfer 500 μL of the stock solution into the first tube. Sequentially transfer 500 μL from the preceding tube into the next tube for subsequent dilutions. Designate the last tube as the blank, do not transfer any solution into it. Aliquot the stock and store at -20 ° C. Use within two weeks and avoid repeated freeze-thaw cycles. Serially diluted standards should be prepared immediately, prior to use.
3. Biotinylated Detection Ab working solution: Calculate the required volume (100 μL/well) plus a slight excess. Centrifuge the Concentrated Biotinylated Detection Antibody (100x) at 800 x g for 1 minute. Dilute to 1x with Biotinylated Detection Ab Diluent at a ratio of 1:99. Prepare the working solution immediately, prior to use.
4. HRP Conjugate Working Solution: Calculate the required volume (100 μL/well) plus a slight excess. Centrifuge the Concentrated HRP Conjugate (100x) at 800 x g for 1 minute. Centrifuge the Concentrated HRP Conjugate (100x) at 800 x g for 1 minute. Prepare the working solution immediately, prior to use.
5. Wash Buffer: Prepare the working Wash Buffer by diluting 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water yielding 750 mL. Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved. For same day use only.

Aufbereitung der Proben:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturation may occur in these samples, leading to false results. Samples should therefore be stored for a short period at 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thaw cycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged to remove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determine compatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibility of 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 the range of the standard curve, the optimal sample dilution for the particular experiment has to be determined.

Testpräzision: Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse 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 Mouse HMGB-1 were tested on 3 different plates, 20 replicates in each plate.
Both intra-CV and inter-CV are < 10 %.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung:

1. For unopened kit: All reagents should be stored according to the labels on the vials, so they are stable up to 12 months after receipt of the kit. The Reference Standard, Biotinylated Detection Antibody, HRP Conjugate and the 96-well stripe plate should be stored at -20 °C upon receipt while the other reagents should be stored at 4 °C.
2. For used kit: When the kit is used, the remaining reagents 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 zip-seal the foil pouch.

Haltbarkeit: 12 months

Referenzen für HMGB1 ELISA Kit (ABIN6962899)

Liu, Xie, Wang, Chen, Han, Wang, Gui, Wu: "JAK2/STAT1-mediated HMGB1 translocation increases inflammation and cell death in a ventilator-induced lung injury model." in: Laboratory investigation; a journal of technical methods and pathology, Vol. 99, Issue 12, pp. 1810-1821, (2020) (PubMed).

Le, Wang, Zhou, Xiong, Tian, Yang, Gai, Sun: "Cigarette smoke-induced HMGB1 translocation and release contribute to migration and NF-κB activation through inducing autophagy in lung macrophages." in: Journal of cellular and molecular medicine, Vol. 24, Issue 2, pp. 1319-1331, (2020) (PubMed).

Ma, Qin, Guo: "MiR-181-5p protects mice from sepsis via repressing HMGB1 in an experimental model." in: European review for medical and pharmacological sciences, Vol. 24, Issue 18, pp. 9712-9720, (2020) (PubMed).

Huang, Hou, Chuan, Wei, Wang, Yang, Ru: "miR-129-5p alleviates LPS-induced acute kidney injury via targeting HMGB1/TLRs/NF-kappaB pathway." in: International immunopharmacology, Vol. 89, Issue Pt A, pp. 107016, (2020) (PubMed).

Liu, Yu, Chen, Sun, Li, Huang, Nie, Wang, Zhang, Gong, Ren: "Hesperidin ameliorates lipopolysaccharide-induced acute lung injury in mice by inhibiting HMGB1 release." in: International immunopharmacology, Vol. 25, Issue 2, pp. 370-6, (2015) (PubMed).

Details zu HMGB1

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

Andere Bezeichnung: High Mobility Group Protein B1

Hintergrund: Target Synonym: HMGB1,HMG1,High mobility group protein B1,High mobility group protein 1 (HMG-1),Hmg-1

Gen-ID: 15289

UniProt: P63158

Pathways: p53 Signalweg, 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