兔血清一氧化氮ELISA原理,兔子NOELISA试剂盒实验步骤

兔血清一氧化氮ELISA原理，兔子NO/ELISA试剂盒实验步骤

Test specimen requirements 1. The specimen is extracted as soon as possible, after extraction, according to the literature should be as soon as possible after extraction experiments. If you can't

Test immediately, can put in the specimen - 20 ℃ to save, but should avoid repeated freezing and thawing 2. Cannot detect samples containing NaN3, because NaN3 inhibition of horseradish peroxidase (HRP) activity. steps

1. Standard dilution: this kit provides the original times standard, the user can be diluted in the small tube according to the following chart Interpretation.

16 standard 150 mu mu g/L 5 L of the original standard to join 150 mu L standard samples times diluent 8 standard 150 mu mu g/L 4 L standard to join 150 mu 5 L standard diluent 4 mu g/L 3 standard 150 mu L 4 standard to join 150 mu L standard diluent 2 mu g/L no. 2 standard 150 mu L 3 standard to join 150 mu L standard diluent 1.0 mu standard 150 g/L 1 2 standard to join 150 mu mu L L standard diluent

2. Add the sample: a blank hole (ck hole without samples and enzyme reagent, the rest of the same step operation), standard hole,

Sample under test. On the enzyme standard package is board standard accurate sample plus 50 mu, l under test sample in the hole and 40 mu l sample diluent,

And then add to test samples of 10 mu l (final sample dilution degree for 5 times). Plus sample will be the sample and the enzyme label plate at the bottom of the hole, do

Don't touch the wall of hole quantity, gently shaking blending.

3. Incubate: use a rear seal plate membrane seal plate temperature of 37 ℃) for 30 minutes. 4. Liquor: 30 times will be concentrated washing liquid reserve 30 times diluted with distilled water

5. Washing: carefully remove the seal plate membrane, abandon to liquid, dry, every hole filled with liquid detergent. Let stand for 30 seconds to refuse, so Repeat 5 times, pat dry.

6. Add enzyme: 50 mu, l per hole plus enzyme standard reagent blank except for hole. 7. Incubate: operation with 3. With 5:8. Washing operation.

9. Color: each Kong Xian adding chromogenic agent A50 mu l, add the chromogenic agent B50 mu l, gently shake blending, 37 ℃ avoid light color 15 minutes.

10. Termination: 50 mu, l per hole and end termination reaction (the blue turn yellow).

11. Determination: the zero blank air conditioning, 450 nm wavelength in order to measure the absorbance (OD value) of each hole.

Determination shall be terminated in Canada

Method a) used to detect the unknown antigen double antibody clip art:

1. The package is: use a 0.05 M PH9 牰 carbonate package is buffer antibodies will be diluted to the protein content of 1 ~ 10 mu g/ml. In each add 0.1 ml, the reaction of polystyrene board hole 4 ℃ for the night. The next day, he went to hole solution, wash with washing buffer 3 times, each time 3 minutes. (hereinafter referred to as catharsis, hereinafter the same).

2. Add: add certain diluted influenza virus samples of 0.1 ml in the above has been the response of pore package, incubated with the 37 ℃ for 1 hour. Then wash. (blank hole at the same time, the bore negative control and positive control).

3. Add enzyme mark antibody: in each reaction in the hole, a fresh diluted enzyme label antibody (after titration of dilution degrees) 0.1 ml. 37 ℃ 0.5 ~ 1 h incubation and washing.

4. Add chromogenic substrate liquid: to add temporary reaction hole configuration of TMB substrate solution 0.1 ml, 37 ℃ for 10 ~ 30 minutes.

5. Termination reaction: add 2 m sulfuric acid in the reaction hole 0.05 ml.

6. Results: on a white background, direct observation with the naked eye: hole the deeper the color reaction, the stronger the degree of positive, negative reaction to colorless or very shallow, on the basis of assumes the color depth, expressed as a \"+\" and \"-\". Also measurable value: o. D on ELISA detector, at 450 nm (if to ABTS color, the 410 - nm), bore to ck after zero measuring the o. D value, if is greater than the negative control stipulated by the OD value of 2.1 times, is positive.

Method 2) used to detect the unknown antibody indirect method:

With package is buffer known antigens will be diluted to 1 ~ 10 mu g/ml, each hole with 0.1 ml, 4 ℃ for the night. The next day washing three times. And must be diluted influenza virus antibody (unknown) 0.1 ml sample in the above has been the response of pore package, 37 ℃ for 1 hour incubation, washing. (at the same time, negative and positive hole blank contrast) in response to hole, a fresh diluted enzyme label the second antibody (anti antibodies) 0.1 ml, 37 ℃ for 30 to 60 minutes incubation, washing, finally with the DDW washing again. The remaining steps with \"double

antibody clip art\" 4, 5, and 6.兔血清一氧化氮ELISA原理，兔子NO/ELISA试剂盒实验步骤

Elisa method is a new technology in the immune diagnosis, has been successfully applied to a variety of infectious diseases and parasitic diseases caused by pathogenic microorganisms and noncommunicable diseases diagnosis. Also has been applied in macromolecular antigen and quantitative evaluation of the small molecule antigens,

according to results have been used, think elisa method is sensitive, specific, simple, fast, stable and easy automation,

etc. Check is not only suitable for clinical specimens, and because a day can check hundreds or even thousands of specimens, therefore, also suitable for serum epidemiology investigation. This law not only can be used to determine antibody, but also can be used for the determination of circulating antigen in body fluids, so it is also a good method of early diagnosis. So the application of elisa method in biomedical fields growing can be summarized in four aspects: 1. The immune enzyme dyeing all kinds of ingredients of localization in the cell. 2. The study of enzyme synthesis of antibodies. 3. Show traces of immune precipitation reaction.

4. The quantitative detection of fluid composition of antigen or antibody.兔血清一氧化氮ELISA原理，兔子

NO/ELISA试剂盒实验步骤

Bring your own things

Enzyme standard instrument (450 nm)

Washer and high precision and spear: 0.5-10 ul, 2-20 ul, 20-200 - ul, ul 200-1000 37 ℃ thermostat Operating considerations

Kit kept in 2-8 ℃, balance for 20 minutes at room temperature before use. Concentrated washing liquid will be taken from the refrigerator, crystallization, which belongs to the normal phenomenon, water bath heating crystallize completely dissolved before use.

Experiments of lath should immediately back into the self-styled bag, seal (low temperature drying).

Number concentration of 0 S0 standard can be seen as negative control or blank. According to the manual operation when samples have been diluted 5 times, the final result actual concentration multiplied by five is the sample.