技术文献

甘薯(Sweet Potato)隐潜病毒(CLV) ELISA检测试剂盒
阅读次数:510 发布时间:2021/5/14 21:20:43
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 Intended use

This CLV ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures.The S Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of CLV in the sample, this CLV ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a cutoff value. The existence or not of CLV in the samples is then determined by comparing the O.D. of the samples to the CUT OFF.

Sample collection and storages

1. Can’t detect the samples which contain NaN3, because NaN3 inhibits HRP activity of the horseradish peroxidase.

2.  Extract as soon as possible after Specimen collection, Extracted according to the relevant literature.

Cell culture supernates and Sweet Potata exact fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw.

Materials required but not supplied

1.  Standard microplate reader(450nm)

2.  Precision pipettes and Disposable pipette tips.

3.  37 ℃ incubator

Precautions

1.  Do not substitute reagents from one kit to another. Standard, conjugate and microplates are matched for optimal performance. Use only the reagents supplied by manufacturer.

2.  Do not remove microplate from the storage bag until needed. Unused strips should be stored at 2-8°C in their pouch with the desiccant provided.

3.  Mix all reagents before using.

Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25°C)

 

 

 

Materials supplied

Name

96 determinations

48 determinations

Microelisa stripplate

12*8strips

12*4strips

Negative control

0.5ml

0.5ml

Positive control

0.5ml

0.5ml

HRP-Conjugate reagent

10.0ml

5.0ml

20X Wash solution

25ml

15ml

Sample Diluent

6.0ml

3.0ml

Chromogen Solution A

6.0ml

3.0ml

Chromogen Solution B

6.0ml

3.0ml

S Solution

6.0ml

3.0ml

Closure plate membrane

2

2

User manual

1

1

Sealed bags

1

1

Reagent preparation

20×wash solution:Dilute with Distilled or deionized water 1:20.

Assay procedure

1.  Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.

2.  Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.

3.  Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing sample well; Blank well doesnt add anyting.

4.  Add 10l of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C. 

5.  Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.

6.  Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.

7.  Add 50μl S Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not

appear uniform, gently tap the plate to ensure thorough mixing.

8.  Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.

 

 

 

 

 

 

Determine the result

1.  Test validity: the average of Positive control well≥1.00; the average of Negative control well ≤0.15.

2.  Calculate Critical (CUT OFF): Critical= the average of Negative control well + 0.15.

    Negative Result: sample OD< Calculate Critical (CUT OFF) is Negative.

    Positive Result: sample OD≥ Calculate Critical (CUT OFF) is Positive.

Storage and validity

Storage:  2-8.

validity six months.

原创作者:上海瑞番生物科技有限公司

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