Human BTA ELISA Test kit

Human BTA ELISA Kit

FOR RESEARCH USE ONLY

Assay range:0.5U/ml - 16U/ml

96 determinations

Purpose

This kit allows for the determination of BTA concentrations in Human serum, cell culture supernates and other biological fluids

Principle of the assay

The kit assay Human BTA level in the sample, use Purified Human BTA antibody to coat microtiter plate wells, make solid-phase antibody, then add BTA to wells, Combined BTA antibody which With HRP labeled, become antibody - antigen - enzyme-antibody complex, after washing Completely, Add TMB substrate solution,TMB substrate becomes blue color At HRP enzyme-catalyzed, reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. The concentration of Human BTA in the samples is then determined by comparing the O.D. of the

samples to the standard curve.

Materials provided with the kit

Specimen requirements

1. extract as soon as possible after Specimen collection,and according to the relevant 2literature, and should be experiment as soon as possible after the extraction. If it can’t,specimen can be kept in -20 ℃ to preserve, Avoid repeated freeze-thaw cycles.

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

Assay procedure

Dilute and add sample:Dilute Original density Standard as follow table:

1.Add sample: Set blank wells separately (blank comparison wells don’t add sample and HRP-Conjugate reagent, other each step operation is same). testing sample well. add Sample dilution 40μl to testing sample well, then add testing sample 10μl (sample final dilution is 5-fold), add sample to wells , don’t touch the well wall as far as possible, and Gently mix.

2. Incubate: After closing plate with Closure plate membrane , incubate for 30 min at 37℃.

3. Configurate liquid: 30-fold(or 20-fold) wash solution diluted 30-fold (or 20-fold) with distilled water and reserve.

4. Washing: Uncover Closure plate membrane, discard Liquid, dry by swing, add washing buffer to every well, still for 30s then drain, repeat 5 times, dry by pat.

5. Add enzyme: Add HRP-Conjugate reagent 50μl to each well, except blank well.

6. Incubate: Operation with 3.

7. Washing: Operation with 5.

8. Color: Add Chromogen Solution A 50ul and Chromogen Solution B 50ul to each well, evade the light preservation for 10 min at 37℃

9. Stop the reaction: Add Stop Solution50μl to each well, Stop the reaction(the blue color

change to yellow color).

10. Assay: take blank well as zero , Read absorbance at 450nm after Adding Stop Solution and within 15min.

Rebecca Yan