Elabscience VB6(Vitamin B6) Elisa Kit

Test principle 
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with VB6. During the reaction, VB6 in samples or Standard competes with a fixed amount of VB6 on the solid phase supporter for sites on the Biotinylated Detection Ab specific to VB6. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are 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 stop solution and the color change is measured spectrophotometrically at a wavelength of 450 ± 2 nm. The concentration of VB6 in the samples is then determined by comparing the OD of the samples to the standard curve.

Kit components & Storage 
An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the items separately according to the following conditions once the kit is received.

Sample collection 
Serum: Allow samples to clot for 1 hour at room temperature or overnight at 2-8℃before centrifugation for 20 min at 1000×g at 2-8℃. Collect the supernatant to carry out the assay. 
Plasma: Collect plasma using EDTA-Na2 as an anticoagulant. Centrifuge samples for 15 min at 1000×g at 2-8℃ within 30 min of collection. Collect the supernatant to carry out the assay.

Calculation of results 
Average the duplicate readings for each standard and samples. Plot a four parameter logistic curve on log-log graph paper, with standard concentration on the x-axis and OD values on the y-axis. If the OD of the sample under the lowest limit of the standard curve, you should re-test it with an appropriate dilution. The actual concentration is the calculated concentration multiplied by the dilution factor.

Dilution method
Please predict the concentration range of samples in advance, and determine the dilution ratio through preliminary experiments or technical support recommendations.