(NADPH)ELISA

Superiority:

Interleukin receptor 17 r test principle:
Interleukin receptor 17 r is solid clip art experiment enzyme-linked immunosorbent (ELISA). The known material under test standard, the concentration of the unknown sample concentration while adding the microporous enzyme label plate for testing.First the material under test and biotin labeled antibody incubate at the same time.After washing, add avidin tagged HRP.Again after incubate and washing, the enzyme that removes not combine, then add in the substrate of A and B, and the enzyme combination at the same time.Color is produced.The depth of the color and the material under test in the sample concentration proportion relationship.


Interleukin receptor 17 r kit contents and its preparation




Interleukin receptor 17 r own material
1) distilled water.
Washer (2) : 5 ul, 10 ul, 50 ul, 100 ul, ul 200 ul, ul 500, 500.
3) oscillator and magnetic stirrer.


Interleukin receptor 17 r safety
1) avoid direct contact with the termination of fluid and the substrate of A and B.Once exposed to the liquid, please rinse with water as soon as possible.
2) experiments do not eat and drink, smoke or use cosmetics.
3) do not use mouth suck kit in any of the ingredients.


Interleukin receptor 17 r operating points for attention
1) reagents should be stored according to label instructions, return to room temperature before use.Dilute the standard should be discarded after the thin, not saved.
2) experiment without batten shall immediately be returned to the bags, sealed preservation, so as not to spoil.
3) no other reagents should be packaged or cover.Don't mix reagents of different batch number.With good quality before use.
4) the use of disposable suction head in order to avoid cross contamination, absorbs the termination and the substrate of A and B, avoid to use washer with metal parts.
5) using a clean plastic container configuration washing liquid.Before using fully blending of all kinds of ingredients and samples in the kit.
6) washing enzyme label plate should be fully taken when dry, don't the blotting paper directly into the enzyme reaction in water absorption.
7) the substrate should be A volatile, avoid long time to open the lid.The substrate B photosensitive, avoid long time exposed to light.Avoid contact with the hand, poisonous.The experiment should be immediately after the completion of OD value.
8) to join the order of reagents should be consistent, to ensure that all reaction plate Kong Wen) time.
The time specified in the 9) according to the instruction, the amount of charging and order to incubate operation.


Interleukin receptor 17 r sample collection, processing and storage
1) serum - operation to avoid any cell stimulation.Use do not contain pyrogen and endotoxin in vitro.After collecting blood, 1000 x g centrifugal 10 minutes to separate serum and erythrocyte quickly carefully.
2) plasma -- -- -- -- -- the EDTA, citrate and heparin plasma can be used for testing.1000 x g centrifugal 30 minutes to remove particles.
3) on the cell supernatant - 1000 x g 10 minutes of centrifugal to get rid of particle and polymer.
4) tissue homogenate -- -- -- -- -- will organize mashed adding suitable amount of saline water.1000 x g centrifugal for 10 minutes, take the supernatant
5) save -- -- -- -- -- - if the sample is not used immediately, should be kept to be divided into small part - 70 ℃, to avoid repeated freezing.As far as possible not to use hemolysis or hyperlipidemia blood.If a large number of particles in serum, centrifugal or filtering before testing.Don't in 37 ℃ or higher temperature heating thawing.Should thaw at room temperature and make sure the sample evenly fully thawed.


Interleukin receptor 17 r reagent preparation
1) standard: standard series of dilution shall be prepared in the experiment, cannot store.The standard of oscillation and mixed before dilution.
2) washing buffer (50 x) dilution: distilled water diluted 50 times.


Interleukin receptor 17 r steps
1) before use, all reagents thoroughly incorporated.Don't make the liquid produces large amounts of foam, so as not to add sample with lots of bubbles and error on the sample.
2) according to the sample under test number plus the number of standard decided the number of panel.Each standard and blank hole advice after hole.To be decided according to the number of each sample, can use the hole after hole as far as possible to do.With specimens were diluent 1:1 diluted 50 ul to join reaction inside the hole.
3) after joining dilution good standard 50 ul joined sample under test in reaction hole, 50 ul in reaction to the hole.Immediately add 50 ul biotin labeled antibody.Cover the diaphragm, oscillation and mixed gently, 37 ℃ incubate for 1 hour.
4) to jilt to hole liquid, each hole filled with detergents, oscillation for 30 seconds, jilt to detergents, pat dry with a blotting paper.Repeat this three times.If use washing machine washing, washing more often.
5) every hole to join 80 ul affinity chain element - HRP enzyme, oscillation and mixed gently, temperature of 37 ℃) for 30 minutes.
6) to jilt to hole liquid, each hole filled with detergents, oscillation for 30 seconds, jilt to detergents, pat dry with a blotting paper.Repeat this three times.If use washing machine washing, washing more often.
7) each hole to join the substrate of A and B 50 ul, oscillation and mixed gently, temperature of 37 ℃) for 10 minutes.Avoid light.
8) remove the enzyme label plate, quickly add 50 ul terminated liquid, to join the determination results immediately after termination of liquid.
9) at 450 nm wavelength determination of OD value of each hole.


Interleukin receptor 17 r limitations
6 standard of the above results for nonlinear, according to the standard curve is unable to obtain accurate results.


Interleukin receptor 17 r kit performance
1. The sensitivity: minimum detectable concentration is less than 1 standard.The linear dilution degrees.Samples of the linear regression correlation coefficient R and expected concentration value of 0.990.
2. Specificity: no reaction with other cytokines.
3. The repeatability, coefficient of variation between plate and plate is less than 10%.


Interleukin receptor 17 r and analysis results
1, instrument value: 450 nm long wavelengths, enzyme mark on the meter reads the OD value of each hole
2, absorbance OD value of the vertical (Y), the corresponding material under test, the abscissa denotes the standard concentration (X), do the corresponding curve, according to the sample of the material under test content OD value by standard curve figure out corresponding concentrations.