Intended use
This immunoassay kit allows for the in vitro quantitative determination of General VB12 concentrations in serum, Plasma, tissue homogenates and Cell culture supernates and Other biological fluids.
Test principle
The ELISA is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to General VB12, During the reaction, General VB12 in the sample or standard competes with a fixed amount of biotin-labeled General VB12 for sites on a pre-coated Monoclonal antibody specific to General VB12. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of General VB12 in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Precision

Intra-assay Precision (Precision within an assay):Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Intra-Assay CV: 4.3%

Inter-assay Precision (Precision between assays):Three samples of known concentration were tested in five separate assays to assess inter-assay precision.

Inter-Assay CV: 7.6%

Recovery

Recovery was determined by spiking various levels of Vitamin B12 into serum and plasma.

Recovery was determined by spiking various levels of VB12 into serum and plasma .

Sample Type	Average(%)	Recovery Range(%)
Serum	99	95-103
Plasma	98	97-102

 

 

 

 

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Vitamin B12 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1：2	1:4	1:8	1:16
serum(n=5)	86-98%	104-117%	104-116%	113-122%
EDTA plasma(n=5)	100-111%	103-113%	109-119%	99-110%
heparin plasma(n=5)	102-111%	91-100%	93-105%	83-92%

 

General Annotation

 

Function:

It is normally involved in the metabolism of every cell of the human body, especially affecting DNA synthesis and regulation, but also fatty acid synthesis and energy production.

 

Location:

Vitamin B12, vitamin B12 or vitamin B-12, also called cobalamin, is a water-soluble vitamin with a key role in the normal functioning of the brain and nervous system, and for the formation of blood. It is one of the eight B vitamins.

 

 

Sequence:C63H88CoN1 | 4O14P

 

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