Vitamin D-binding Protein, Human, BioAssay(TM) ELISA Kit (DBP, VDB, Gc-globulin, Group-specific Component, GC)

Vitamin D-binding protein (DBP, VDBP), also called group-specific component (Gc) and macrophage-activating factor (GcMAF/DBP-MAF), is 52 to 58kD plasma glycoprotein with many functions, such as transport of vitamin D metabolites, control of bone development, binding of fatty acids, sequestration of actin, and modulating immune and inflammatory responses. DBP is synthesized predominantly by hepatic parenchymal cells and detected in plasma, cerebrospinal fluid, seminal fluid, saliva and breast milk. The exploitation of the unique properties of DBP could enable the development of important therapeutic agents such as vitamin D-associated conditions, actin sequestering in trauma and inflammation, chronic obstructive pulmonary disease, osteopetrosis, cancer therapy and immune modulation by macrophage activation. The DBP molecule is therefore an ideal candidate molecule for further investigation by biotechnology-based companies seeking a platform from which to pursue new therapeutic options.\n\nIntended Use:\nHuman VDBP ELISA kit is to be used for the in vitro quantitative determination of human VDBP in human serum, human plasma, cell lysate and buffered solution. The assay will recognize native human VDBP. 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 VDBP was calculated to be 2.8ng/ml, by subtracting two standard deviations from the mean of 12 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: human serum albumin, transferrin, IgG, alpha-fetoprotein(AFP), fibrinogen, plasminogen, Hemoglobin.\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 Fibrinogen. Samples are pippetted into these wells. Nonbound fibrinogen and other components of the sample should be removed by washing, then biotin- conjugated monoclonal antibody specific to fibrinogen added. In order to quantitatively determine the amount of fibrinogen 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 VDBP.\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 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

V2130-10M

Size

96Tests

References

1. Gomme, P.T. and Bertolini, J. Trends Biotechnol. (2004) vol.22(7):340-345. 2. Svasti, J. et al., Biochemistry (1979) vol.18:pp.1611-1617. 3. White, P. and Cooke, N., Trends Endocrinol. Metab. (2000) vol.11:pp. 320-327.