Sample
Serum, plasma, cell culture supernatant and other biological samples
Intended Use
In vitro quantitative determination of HMGB1 concentrations in serum, plasma, cell culture supernatant and other biological samples.
Contents of Kit
1. ELISA Microplate(Dismountable) : 96T(8×12). Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C.
2. Lyophilized Standard: 2vial. Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C.
3. Biotin-labeled Antibody (Concentrated, 100×): 120 ul. 2-8°C (Avoid Direct Light)
4. HRP-Streptavidin Conjugate (SABC, 100×): 120 ul. 2-8°C (Avoid Direct Light)
5. TMB Substrate: 10 ml. 2-8°C (Avoid Direct Light)
6. Sample Dilution Buffer: 20 ml
7. Antibody Dilution Buffer: 10 ml
8. SABC Dilution Buffer: 10 ml
9. Stop Solution: 10 ml
10. Wash Buffer (25×): 30 ml
11. Plate Sealer: 5
Note: The liquid reagent bottle contains slightly more reagent than indicated on the label. Please use pipette accurately measure and do proportional dilution.
Storage
Store at 2-8°C (for sealed box), please do not freeze! See kit label for expiry date.
StabilityPerform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
Precision
Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
Detection Range
31.25-2000pg/ml
Standard Curve
This product has been tested by Quality Control Department and meets performance specifications mentioned in the manual. (The humidity in the laboratory is 20%-60%, and the temperature is 18°C - 25°C. TMB was balanced to 37°C before color development, and incubated at 37°C for 15 minutes in the dark after adding the enzyme label plate holes.)
The following assay data are provided for reference, since experimental environment and operation are different. The establishment of standard curve depends on your own assay.

Citations
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Yamada, S; Inoue, K; et al. High mobility group protein 1 (HMGB1) quantified by ELISA with a monoclonal antibody that does not cross-react with HMGB2. CLINICAL CHEMISTRY 49:1535-1537(2003).
Ito, I; Mitsuoka, N; et al. Conformational difference in HMGB1 proteins of human neutrophils and lymphocytes revealed by epitope mapping of a monoclonal antibody. JOURNAL OF BIOCHEMISTRY 136:155-162(2004).