Free Testosterone( Free Tes ) ELISA Test Kit

For in vitro diagnostic and professional use only.

Drug Names

Generic Name: Ferr Testosterone ELISA Test Kit (Serum/Plasma)

INTENDED USE

This kit is a quantitative detection of Ferr Testosterone (Ferr Tes) in human serum/plasma in vitro. The kit is suitable for clinical screening and diagnosis.

Product details

PRINCIPLE

This kit uses the Competitive-ELISA principle. The microtiter plate strips has been pre-coated with Coating antibody for Testo. Standards or samples containing Testo and biotin labeled Testo are added to the plate, A competitive inhibition reaction is launched between labeled Testo and free Testo (Standards or samples) with the solid phase antibody specific to Testo. After that, Streptavidin-Horseradish Peroxidase(SA-HRP) is added to form a sandwich complex of solid phase antibody-biotin labeled antigen-SA-HRP. And then, TMB substrate solution is added to all wells and incubated. An enzyme-catalyzed reaction generates a blue color in the solution, thereafter, stop solution is added to stop the substrate reaction and the color turns yellow. The yellow solution is read at a wavelength of 450nm. The concentration of Testo in the samples is then calculated from the ODvalue by establishing a standard curve.

TEST PROCEDURE

1. Add the Standard working solution to the first two columns: Each concentration of the solution is added in duplicate, to one well each, side by side (50μL for each well). Add the samples to the other wells (50 μL for each well).

2. Add 50μL of Detection Reagent A working solution to each well. Cover with the plate sealer. Incubate for 60 min at 37℃.

3. Discard the liquid from each well, add 350μL of wash buffer to each well. Soak for 30sec and decant the solution from each well and pat it dry against clean absorbent paper. Repeat this wash step 3 times in total.

4. Add 100μL of Detection Reagent B working solution to each well. Cover with the plate sealer. Incubate for 30 min at 37℃.

5. Turn on the microplate reader for preheating.

6. Discard the liquid from each well, repeat the washing process of step 3 for 5 times.

7. Add 90μL of TMB Reagent to each well. Cover with a new plate sealer. Incubate for 10-20 min at 37℃. Protect the plate from light.

8. Add50 μLofStopSolution to each well. 9. The absorbance OD value of each well is measured at 450nm.

Important notes

• The kit takes out from the refrigeration environment should be balanced 15-30 minutes in the room temperature then use, ELISA plates coated if has not use up after opened, the plate should be stored in Sealed bag.
• washing buffer will Crystallization separation, it can be heated the water helps dissolve when

dilute . Washing does not affect the result.

• add Sample with sampler Each step, And proofread its accuracy frequently, avoids the experimental error. add sample within 5 min, if the number of sample is much , recommend to use Volley .
• Please make specification curve when you assay, had better make duplicate well, if in the sample the testing material content is excessively high (The sample OD is bigger than the first standard well OD ),please use Sample dilution to dilute certain multiple (n times),then assay. Please multiply total Dilution Times when calculate(×n×5).
• Closure plate membrane only limits the disposable use, in order to avoid the overlapping pollution.
• The substrate please evade the light preservation.
• Please according to use instruction strictly, The test result determination must take the microtiter plate reader as a standard .
• All samples, washing buffer and each kind of reject should according to infective material process.
• This reagent which different batch number component do not mix.
• If it’s different form English instruction, take English instruction as the standard.

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.