Vitamin E Elisa Test Kit

Drug Names

Generic Name:Vitamin E Elisa Test Kit

Purpose

This kit a lows for the determination of VE concentrations in sample.

Principle of the assay

The kit use ELISA competition to assay VE level in the sample,use Purified VE antibody to coat microtiter plate we ls, make solid-phase antibody, add VE and antibody which With HRP labeled VE to coated microtiter wels,make them Combine competitive ,after washing Completely, Add TMB substrate solution . the depth of color and the VE of sample were positively correlated, measure the optical densit (OD) at 450 nm with microtiter plate reader, calculate VE concentration by standard curve.

Specimen requirements

10 Closure plate membrane 2 1

1．Specimen process:

(1) Water samples repeated freeze-thaw cycles three times after extraction. then filter with glass fiber, reference.

(2) Tissue Samples with n-butanol: methanol: water (5:25:70 V: V: V) extraction, or extract according to related literatures. experiment as soon as possible after the extraction. If it can not be tested immediately, specimen can be kept in-20 ℃, reference.

2. Can’t detect the sample which contain NaN3, because NaN3 inhibits HRP activity .

dilute . Washing does not affect the result.

Assay procedure

• add sample:Set blank wels separately (blank comparison wels don’t add sample and ELISA reagent, other each step operation is same). testing sample wel. add Sample dilution 40μl to testing sample we l which on ELISA plates coated, then add testing sample 10μl (sample final dilute degree is 5 times), add sample to the bottom of ELISA plates coated wel , don’t touch the we l wal as far as possible, and Gently mix.
• add enzyme:Add ELISA reagents 50μl to each wel, except the blank wel. Incubate: After closing plate with Closure plate membrane ,incubate for 30 min at 37℃.
• Configurate liquid: 30 times of Wash Concentrate diluted 30 times with disti led water and reserve.
• washing:Uncover Closure plate membrane, discard Liquid, dry by swing, add washing buffer to every we l, sti l for 30 second then remove, repeat 5 times, dry by pat.
• color:add color reagent A 50μl and color reagent B 50μl to each we l. Gently mix, incubate for 10 min at 37℃.
• Stop the reaction:Add Stop Solution50μl to each wel, Stop the reaction(the blue color change to yelow color Immediately).
• assay:take blank wel as zero , measure the optical densit (OD) at 450 nm after Adding Stop Solution and within 15min..

Storage and stability

• Store at 2-8℃.

• Seal and return unused reagents to 2-8℃, under which conditions the stability will be retained for 2 months, or until the labeled expiry date, whichever is earlier.