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小鼠仙台病毒(Sendai)ELISA试剂盒使用说明书

Mouse Sendai Virus ELISA Kit

USE INSTRUCTION

FOR RESEARCH USE ONLY

Drug Names

Generic Name：Mouse Sendai Virus ELISA Kit.

Purpose：

This kit allows for the determination of Sendai Virus concentrations in mouse serum, and other related samples.

Principle of the assay

The kit use ELISA to assay Sendai Virus in the sample, use Purified Sendai Virus antibody to coat microtitration wells, make solid-phase antibody, which combined with Sendai Virus, after washing and removing non-combinative antibody and other components , Combined antibody which With HRP labeled become antibody - antigen - enzyme-antibody complex, after washing Completely, Add TMB substrate solution and color develops, TMB becomes blue color At HRP enzyme-catalyzed, And at the effect of acid the color finally become yellow, measure the optical densit (OD) at 450 nm with microtiter plate reader, Compared with the CUTOFF value, according to this to judge mouse Sendai Virus exist in the sample or not.

Materials provided with the kit

Specimen requirements

1.      Specimen process：（1）serum or blood plasma  can be assay directly.

                     （2）other samples: Prepared according to relevant literature.

2. experiment as soon as possible after preparing. If it can not be tested immediately,specimen can be kept in -20 ℃ a month to preserve, Avoid repeated freeze-thaw cycles.

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

Assay procedure

1.        Number: to sample correspond microtitration well and Number Sequence, each plate should be set feminine comparison 2 wells, masculine comparison 2 wells, blank comparison 1 well(don’t add sample and HRP-Conjugate reagent to blank comparison well, other each step the operation is same).

2.        add sample：separately add feminine comparison and masculine comparison(Standard) 50μl to the feminine and masculine well . add Sample dilution 40μl to testing sample well, then add testing sample 10μl. add sample to the bottom of ELISA plates coated well , don’t touch the well wall as far as possible, and Gently mix.

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

4.       Configurate liquid: add distilled water to 20 times of Wash solution until 1000ml,and reserve.

5.        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.

6.        add enzyme：Add ELISA reagents 50μl(or drop ) to each well, except the blank well.

7.        incubate：Operation with 3.

8.        washing：Operation with 5.

9.        color：add color reagent A 50μl(or drop ) and color reagent B 50μl(or drop ) to each well. Gently mix, incubate for 15 min at 37℃.

10.    Stop the reaction：Add Stop Solution50μl to each well, Stop the reaction(the blue color change to yellow color Immediately).

11.    assay：take blank well as zero , measure the optical densit (OD) at 450 nm after Adding Stop Solution and within 15min.

Determine the result

Test validity: the average of masculine comparison well≥1.00; the average of feminine comparison well ≤0.15.

Calculate Critical(CUT OFF) : Critical= the average of masculine comparison well + 0.20.

feminine determination: sample OD< Calculate Critical(CUT OFF) is MHV feminine.

Masculine determination: ample OD≥ Calculate Critical(CUT OFF) is MHV Masculine.

Important notes

1． Please according to use instruction strictly, This reagent which different batch number component don’t mix.

2． 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.

3.      washing buffer will Crystallization separation, it can be heated the water helps dissolve when dilute . Washing does not affect the result.

4． Closure plate membrane only limits the disposable use, in order to avoid the overlapping pollution

5．The substrate please evade the light preservation.

6．The test result determination must take the microtiter plate reader as a standard, when use dual-wavelength to assay, Reference wavelength is 630nm.

7．All samples, washing buffer and each kind of reject should according to infective material process. Stopp Solution is 2M sulphuric acid. You must pay attention to safe when use .

8. If it’s different form English instruction, take English instruction as the standard.

Package size

96 determinations.

Storage and validity

1．Storage： 2-8℃.

2．validity： six months.

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