Vitamin B12 Microtiter Plate Assay Kit


The Eagle Biosciences Vitamin B12 is a microtiter plate test kit based on a microbiological assay which measures the total Vitamin B12 content in serum. The test kit contains all required reagents, e.g. standard, medium and microtiter plate coated with a specific microorganism, sufficient for 96 determinations including standard curves. An ELISA reader is required for evaluation of the Vitamin B12 content.  The Vitamin B12 Assay Kit is for research use only. 

SKU: KIFR012 Categories: , ,

Vitamin B12 Microtiter Plate Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 6 ng/L
Dynamic Range: 6 – 54 ng/L
Incubation Time: 48 hours
Sample Type: Serum
Sample Size: 100 µL

Product manufactured in Germany by Immundiagnostik

Additional Information

Assay Background

Vitamin B12 (Cobalamin), a collective term for a group of variously substituted corrinoid with cobalt as the central atom, is found free and also protein-bound in food. The protein-bound form is degraded by pancreatic protease, releasing free B12 which binds to intrinsic factor, a protein secreted by gastric parietal cells of the stomach mucosa. The Cobalamin-Intrinsic-Factor-Complex is bound to mucous membrane cells of the stomach in the ileum and absorbed by the cells. In the case of high doses, a diffusion of the complex also takes place. Vitamin B12 is bound to the protein Transcobalamin II (TC-II) within the cells. TC-II serves as a transport protein for Vitamin B12 in the circulation system. Vitamin B12 is involved in the metabolism process as a co-enzyme and plays an important role in the formation of the blood, the development of the nervous system and the regeneration of the mucous membranes. In addition, there is a direct relationship to the formation of folic acid because methylcobalamin is involved in the transfer of methyl groups for the synthesis of methionine from homocysteine.

Assay Principle

The serum is pre-treated and diluted with a buffer mixture, and then samples were transferred in the wells of a microtiter plate [PLATE] coated with Lactobacillusdelbrueckii subsp.lactis.The addition of vitamin B12 in either standards [STD] or samples gives a vitamin B12-dependent growth response until vitamin B12 is consumed. After incubation at 37°C for 44 – 48 hours, the growth of Lactobacillusplantarum is measured turbidimetrically at 610 – 630 nm (alternatively at 540 – 550 nm) in an ELISA-reader and a standard curve is generated from the dilution series. The amount of vitamin B12 is directly proportional to the turbidity.


Product Manual