Vitamin B12, Food BioAssay(TM) ELISA Kit

Vitamin B12 as a trace element belongs to the biologically important chelate formers. The basic unit consists of a corrin ring with cobalt as a central atom. Cobalt is sixfold coordinated by four nitrogen atoms, one cyanide and a dimethylbenzimidazol group. Vitamin B12 forms a stable complex, which is absorbed in the lower part of the small intestine, with the so-called intrinsic factor present in the gastric juice. A lack of vitamin B12 can lead among other things to pernicious anemia. This disease is not generated by an insufficient supply of vitamin B12, but by the absence of intrinsic factor. A pernicious anemia can be treated by a high dosage of vitamin B12.\n\nThe existing detection procedures are mainly microbiological methods, but also HPLC and thin-layer chromatography, all of which are associated with a high amount of time and instrumentation. With the present test kit it is possible, to determine vitamin B12 quantitatively in vitaminated food in a significantly faster way (2.5 to 4 hours inclusive sample pretreatment) compared with a conventional microbiological assay (24 to 48 hours).\n\nIntended Use:\nFor the quantitative determination of Vitamin B12 in food.\n\nSensitivity: 0.3ng/ml\n\nRecovery (spiked samples): ~98%\n\nIncubation Time: 80 min\n\nTest Principle:\nThe United States Biological Vitamin B12 quantitative test is based on the principle of the enzyme linked immunosorbent assay. An antibody directed against vitamin B12 is bound on the surface of a microtiter plate. Vitamin B12 containing samples or standards and a vitamin B12-peroxidase conjugate are given into the wells of the microtiter plate. Enzyme labeled and free vitamin B12 compete for the antibody binding sites. After one hour incubation at room temperature, the wells are washed with diluted washing solution to remove unbound material. A substrate solution is added and incubated for 20 minutes, resulting in the development of a blue color. The color development is inhibited by the addition of a stop solution, and the color turns yellow. The yellow color is measured photometrically at 450nm. The concentration of vitamin B12 is indirectly proportional to the color intensity of the test sample.\n\nKit Components:\nV2126-01A: Microtiter Plate, 1x96 wells each, coated with anti-vitamin B12.\nV2126-01B: Standard 0ng/ml, 1x500ul, ready to use.\nV2126-01C: Standard 0.4ng/ml, 1x500ul, ready to use.\nV2126-01D: Standard 1ng/ml, 1x500ul, ready to use.\nV2126-01E: Standard 4ng/ml, 1x500ul, ready to use.\nV2126-01F: Standard 10ng/ml, 1x500ul, ready to use.\nV2126-01G: Standard 40ng/ml, 1x500ul, ready to use.\nV2126-01H: Vitamin B12 (HRP), 1x6ml ,ready-to-use.\nV2126-01J: Substrate Solution (TMB), 1x15ml, ready to use.\nV2126-01K: Stop Solution (0.5M H2SO4), 1x15ml, ready to use.\nV2126-01L: Sample Diluent, 2x60ml, ready to use.\nV2126-01M: Washing Solution, 10x, 1x30ml \n\nStorage and Stability:\nStore components at 4 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.\n\nSensitivity: 0.3ng/ml\n\nRecovery (spiked samples): ~98%\n\nIncubation Time: 80 min\n\nIntra-assay Precision: 3%

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SPECIFICATIONS

Catalog Number

V2126-01

Size

1Kit

Applications

ELISA

References

1. Thompson, M.T. et al; J. Biol. Chem. 184, 175 (1950). 2. Reichert, N., Rubach, K.; Dt. Lebensmittel-Rundschau 86/10 (1990). 3. Miller A., Slingerland D.W, Hall C.A., Chu R.C.; Am J Hematol. 1998 59:42-5. 4. Segal, R., Baumoehl, Y., Elkayam, O., Levartovsky, D., Litinsky, I., Paran, D. , Wigler, I., Habot, B., Leibovitz, A., Sela, B.A., Caspi, D.; Rheumatol Int. 2004 24:14-9. Epub 2003 Apr 29. 5. Clarke, R., Smith, A.D., Jobst, K.A., Refsum, H., Sutton, L., Ueland, P.M.; ArchNeurol. 1998 55:1449-55. 6. Bernard, M.A., Nakonezny, P.A., Kashner, T.M.; J Am Geriatr Soc. 1998 46:1199-206.