DBI, Human, BioAssay(TM) ELISA Kit (Acyl-CoA-binding Protein, ACBP, Diazepam-binding Inhibitor, Endozepine, EP)

Diazepam Binding Inhibitor/Acyl-coA binding protein (DBI, ACBP) is a highly conserved 10kD polypeptide which is expressed in various species range from yeast to mammals. As an inverse agonist for benzodiazepine receptors, DBI (ACBP) down-regulates inhibitory effects of GABA. It also has potential to induce anxiety. Found in central and peripheral tissues, DBI (ACBP) also participates in metabolism of steroids, which has been known to partially modify GABA receptor function in the CNS. In peripheral tissues, DBI(ACBP) plays regulatory roles in steroidogenesis. DBI (ACBP) levels have been reported to be decreased in the cerebrospinal fluid obtained from patients with Alzheimer disease.\n\nIntended Use:\nDiazepam Binding Inhibitor/ Acyl-coA binding protein (human DBI, ACBP) ELISA kit is to be used for the in vitro quantitative determination of human DBI(ACBP) in human serum, human plasma, cell lysate and buffered solution. The assay will recognize both native and recombinant human DBI(ACBP). This kit has been configured for research use only and is not to be used in diagnostic procedures.\n\nSensitivity:\nThe minimal detectable dose of human DBI(ACBP) was calculated to be 25pg/ml, by subtracting two standard deviations from the mean of 10 zero standard replicates (ELISA buffer, S0) and intersecting this value with the standard curve obtained in the same calculation.\n\nSpecificity:\nThe following substances were tested and found to have no cross-reactivity: mouse DBI (ACBP) and rat DBI(ACBP).\n\nTest Principle:\nThe design of this assay is based on a sandwich Enzyme-Linked Immunosorbent Assay (ELISA). The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific to human DBI (ACBP). Samples are pippetted into these wells. Nonbound DBI (ACBP) and other components of the sample should be removed by washing, then biotin- conjugated monoclonal antibody specific to DBI (ACBP) added. In order to quantitatively determine the amount of DBI (ACBP) present in the sample, Avidin conjugated to Horseradish Peroxidase (HRP) should be added to each microplate well. The final step, a TMB-substrate solution added to each well. Finally, a sulfuric acid solution is added and the resulting yellow colored product is measured at 450nm. Since the increases in absorbency is directly proportional to the amount of captured DBI (ACBP).\n\nKit Components:\nMicroplate: 1x96 wells\nIncubation buffer: 1x30ml\nWashing buffer, (10x): 1x100ml\nStandard protein: 1 glass vial lyophilized\nStandard/sample dilution buffer: 1x25ml\nSecond antibody: 1 glass vial lyophilized\nAV-HRP, (100x): 1x150ul\nSecondary antibody/AV-HRP dilution buffer: 1x25ml\nSubstrate (TMB): 1x20ml\nStop solution: 1x20ml\n\nStorage and Stability:\nStore components at 4 degrees C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

D3475-02Q

Size

96Tests

References

1. Andoh, T. et al. (2002) J.Biol.Chem. 277, 9655-9660. 2. Arner, E. S. and Holmgren, A. (2000) Eur. J. Biochem. 267, 6102-6109. 3. Lundstrom, J. and Holmgren, A. (1990) J. Biol. Chem. 265, 1994-9120. 4. Nordberg, J. and Arner, E. S. J. (2001) Free Radic. Biol. Med. 31, 1287-1312. 5. Matthews, J. R. et al. (1992) Nucleic Acids Res. 20, 3821-3830. 6. Wei, S. J. (2000) Cancer Res. 60, 6688-6695. 7. Chae, H. Z. (1999) Methods Enzymol. 300, 219-226 IMMUNOBLOT. 8. Hoshino, Y. et al. (2007) Antioxid Redox Signal. 9.689-99.