Porcine CSFV Ag ELISA Kit

Porcine CSFV Ag ELISA Kit

FOR RESEARCH USE ONLY

96 determinations

Purpose

This kit allows for the determination of CSFV Ag concentrations in Porcine

serum, and other biological fluids.

Principle of the assay

The kit assay CSFV Ag level in the sample，use Purified CSFV antibody to

coat microtiter plate wells, make solid-phase antibody, then add CSFV Ag to

wells, Combined With CSFV Ag, after washing and removing non-combinative

antibody and other components ,then Combined CSFV 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. Compared with the

CUTOFF value, according to this to judge CSFV Ag exist in the sample or not.

Materials provided with the kit

1 wash solution 20ml×1bottle 7 Stop Solution 6ml×1 bottle

2 HRP-Conjugate

reagent 6ml×1 bottle 8 Positive control 0.5ml×1 bottle

3 Microelisa stripplate 12well×8strips 9 Negative

control

0.5ml×1bottl

e

4 Sample diluent 6ml×1 bottle 10 Instruction 1

5 Chromogen Solution

A 6ml×1 bottle 11 Closure plate

membrane 2

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6 Chromogen Solution

B 6ml×1 bottle 12 Sealed bags 1

Specimen requirements

1. extract as soon as possible after Specimen collection,and according to the

relevant literature, 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

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 are same).

2.add sample：separately add Positive control and Negative control 50μl to the

Positive and Negative 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: 30-fold (or 20-fold)wash solution diluted 30-fold (or 20-fold)

with distilled water until 600ml,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

IBL International GmbH

Flughafenstra?e 52a 22335 Hamburg DE

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pat.

6.add enzyme：Add HRP-Conjugate reagent 50μlto each well, except the blank

well.

7.incubate：Operation with 3.

8.washing：Operation with 5.

9.color：Add Chromogen Solution A 50ul and Chromogen Solution B to each

well, evade the light preservation 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).

11. assay：take blank well as zero , Read absorbance at 450nm after Adding

Stop Solution and within 15min.

Determine the result

Test validity: the average of Positive control well≥1.00; the average of

Negative control well ≤0.10.

Calculate Critical(CUT OFF) : Critical= the average of Negative control well

+ 0.15.

Negative control: sample OD< Calculate Critical(CUT OFF) is CSFV Ag

Negative control.

Positive control: ample OD≥ Calculate Critical(CUT OFF) is CSFV Ag

Positive control.

Important notes

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Flughafenstra?e 52a 22335 Hamburg DE

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1.Please according to use instruction strictly, Do not mix reagents with those

from other lots.

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 .

Storage and validity

1．Storage： 2-8℃.

2．validity： six months.