Human Epstein-Barr virus capsid antigen IgM (EB VCA IgM) enzyme-linked immunosorbent assay

Drug Name:
Generic name: human EB virus capsid antigen IgM (EB VCA IgM) enzyme-linked immunoassay kit Use:
This kit is a qualitative test for EB virus capsid antigen IgM (EB VCA IgM) in human blood or other related tissues.
Experimental principle:
The kit uses a double antibody sandwich enzyme-linked immunosorbent assay (ELISA) to determine the human EB virus capsid antigen IgM (EB VCA IgM) in the specimen. The microplate was coated with purified anti-EB virus capsid antigen IgM (EB VCA IgM) to prepare a solid phase antibody, which can be combined with the human EB virus capsid antigen IgM (EB VCA IgM) in the sample and washed to remove The unbound antigen and other components are then combined with the HRP-labeled anti-EB virus capsid antigen IgM (EB VCA IgM) to form an antibody-antigen-enzyme-labeled antibody complex, which is thoroughly washed and then primed with TMB. TMB is converted to blue under the catalysis of HRP enzyme and converted to the final yellow color of zui under the action of acid. The absorbance (OD value) was measured at a wavelength of 450 nm using a microplate reader, and compared with the CUTOFF value to determine the presence or absence of the human EB virus capsid antigen IgM (EB VCA IgM) in the specimen.
Kit composition:
1 20 times concentrated washing solution 50ml × 1 bottle 7 Stop solution 6ml × 1 bottle
2 enzyme standard reagent 6ml × 1 bottle 8 positive control 0.5ml × 1 bottle
3 enzyme label coating plate 12 holes × 8 9 negative control 0.5ml × 1 bottle
4 sample dilution 6ml × 1 bottle 10 instructions 1
5 developer A liquid 6ml × 1 bottle 11 sealing film 2 sheets
6 developer B liquid 6ml × 1 bottle 12 sealed bag 1 specimen requirements:
1. Specimen processing: serum and plasma samples can be directly detected
2. The specimens were tested as soon as possible after preparation. If it is not detected in time, the specimen can be stored at -20 °C for one month, but repeated freezing and thawing should be avoided.
3. Samples containing NaN3 could not be detected because NaN3 inhibited horseradish peroxidase (HRP) activity.
Steps:
1. No.: The corresponding micropores of the sample are numbered sequentially. Each plate should have 2 holes for negative control, 2 holes for positive control and 1 well for blank control (no blank sample and enzyme standard reagent for blank control well, the other steps are the same)
2. Loading: 50 μl of negative control and positive control (standard) were added to the negative and positive control wells, respectively. Then, add 40 μl of the sample dilution to the sample well to be tested, and then add 10 μl of the sample to be tested. Add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, and shake it gently.
3. Incubation: The plate was sealed with a sealing film and incubated at 37 ° C for 30 minutes.
4. Liquor: 20 times concentrated washing solution plus distilled water to 1000ml
5. Washing: Carefully remove the sealing film, discard the liquid, dry it, fill each well with the washing solution, let stand for 30 seconds, then discard it, repeat 5 times, and pat dry.
6. Add enzyme: Add 50 μl of enzyme labeling reagent to each well, except for blank wells.
7. Incubation: The operation is the same as 3.
8. Wash: Operate the same as 5.
9. Color development: add 50 μl of developer A to each well, then add 50 μl of developer B, gently shake and mix, and color at 37 ° C for 15 minutes.
10. Termination: 50 μl of stop solution was added to each well to stop the reaction (the blue color turned yellow).
11. Measurement: The absorbance (OD value) of each well was measured in sequence with a blank air conditioner of zero and a wavelength of 450 nm. The measurement should be carried out within 15 minutes after the addition of the stop solution.

The result is judged:
Test validity: mean value of positive control well ≥ 1.00; average value of negative control ≤ 0.15
CUT OFF calculation: critical value = negative control well average + 0.15
Negative judgment: sample OD value < CUT OFF is human EB virus capsid antigen IgM (EB VCA IgM) negative positive judgment: sample OD value ≥ critical value (CUT OFF) is human EB virus capsid antigen IgM (EB VCA IgM) positive considerations
1. The operation is carried out in strict accordance with the instructions. The components of the different batches of this reagent shall not be mixed.
2. The kit should be taken out from the refrigerated environment and should be used at room temperature for 15-30 minutes before use. If the enzyme label is unsealed after opening, the slats should be stored in a sealed bag.
3. Concentrated washing liquid may crystallize out. When diluted, it can be heated and dissolved in a water bath. The washing will not affect the result.
4. The sealing film is intended for single use only to avoid cross-contamination.
5. Please keep the substrate away from light.
6. The test results must be determined by the microplate reader. When using dual-wavelength detection, the reference wavelength is 630nm.
7. All samples, washings and various wastes should be treated as infectious materials. The stop solution is 2M sulfuric acid and must be used safely.
specification:
96 person/box storage conditions and expiration date
1. The kit is stored at: 2-8 °C.
2. Validity: 6 months

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