Intended use
This immunoassay kit allows for the in vitro quantitative determination of General HCY 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 HCY, During the reaction, General HCY in the sample or standard competes with a fixed amount of biotin-labeled General HCY for sites on a pre-coated Monoclonal antibody specific to General HCY. 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 HCY 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: ≤5.0%

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: ≤8.0%

Recovery

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

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

Sample Type	Average(%)	Recovery  Range (%)
Serum	85	79-93
Plasma	92	85-101

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Homocysteine 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)	100-110%	113-123%	97-107%	91-101%
EDTA plasma(n=5)	92-102%	82-92%	105-115%	101-110%
heparin plasma(n=5)	107-117%	93-105%	98-111%	105-114%

 

General Annotation

 

Function:

Homocysteine is a non-protein amino acid with the formula HSCH2CH2CH(NH2)CO2H. It is a homologue of the amino acid cysteine, differing by an additional methylene (-CH2-) group. It is biosynthesized from methionine by the removal of its terminal Cε methyl group. Homocysteine can be recycled into methionine or converted into cysteine with the aid of B-vitamins. While detection of high levels of homocysteine has been linked to cardiovascular disease, lowering homocysteine levels may not improve outcomes.

 

 

Sequence:C4H9NO2S

 

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