Human ELISA kit transfers chemotactic growth factor
This kit is for research use only: serum or plasma
1. Matters needing attention
1. This reagent is an in vitro test reagent and is used within the validity period. The reagent should be regarded as an infectious agent. Reagents with different batch numbers cannot be mixed.
The reagents in the box should be taken out before use. Leave at room temperature for at least 30 minutes,
After the concentrated washing solution crystallizes, incubate at 37 ° C for 15 minutes.
Incubate the human ELISA kit at 37 ° C for 15 minutes after crystallization of the concentrated sample diluent
If the experiment is carried out within 24 hours, the specimen can be stored at 2 ～ 8 ℃. No need for timely experiment, the specimen should be stored at -20 ℃ to avoid repeated freezing and thawing.
After repeatedly cleaning the microwell plate, and buckle the residual liquid in the microwell, otherwise it will reduce the accuracy and cause the false image of absorbance deviation.
After the sample is added, the microwell reaction strip should be shaken slightly to mix the liquid in the well.
The kit is stored at 2 ～ 8 ℃, please do not freeze it, please refer to the label in the box for the expiration date.
2. Preparation before the experiment
1. Before use, remove all reagents in the box and leave at room temperature for at least 30 minutes.
2. Prepare various experimental instruments and materials, such as micro pipettes, pipette tips, medical distilled water, etc.
3. Concentrated lotion and medical distilled water are diluted 1:19 times to become an applied lotion
4. Dilute the concentrated sample diluent and medical distilled water 1: 4 times into the applied sample diluent
5. Dilute the sample with the application sample diluent, dilute the sample according to the volume ratio of 1: 100, for example, 10μl of the sample is added to the 1ml application sample diluent, and mix well to be used. )
3. Operation steps
1. Take out the enzyme labeling plate and set up blank wells. Add 100μl of the standard products to the blank micro-wells according to the order (the blank wells are regarded as No. 0 standards and replaced with medical distilled water)
2. Mark the sample number separately and add 100 μl of the diluted sample to the blank microwell (different samples use different tips).
3. Incubate the microplate at 37 ° C for 30 minutes;
4. Take out the enzyme labeling plate and shake off the liquid in it. After filling all the holes with the application washing liquid, immediately shake off the liquid;
5. After the application washing solution is filled in each well, shake the enzyme plate slightly for 30 seconds, then shake off the application washing solution in the well, and pat the enzyme plate dry on the absorbent paper.
6. Repeat the fifth step 5 times, pat the enzyme plate dry on absorbent paper.
7. Add 100 μl of enzyme-labeled coupling solution to the standard well and sample well.
8. Incubate the 96-well plate at 37 ° C for 30 minutes.
9. Take out the enzyme labeling plate and shake off the liquid. After all the application washing liquid is filled in each well, immediately shake off the liquid.
10. After refilling each well with the washing application solution, shake the enzyme plate slightly at room temperature for 30 seconds, then shake off the application washing solution in the wells, and pat the enzyme plate on the absorbent paper to dry.
11. Repeat the fifth step 5 times, pat the enzyme plate on the absorbent paper to dry.
12. Add 50 μl of substrate A to each well immediately after adding 50 μl of substrate B. Gently shake to mix. (Liquid A and B use different tips to add samples)
13. Incubate the microplate at 37 ° C in the dark for 15 minutes.
14. Add 50 μl of stop solution to each microwell and gently shake to mix.
15. Measure OD at 450nm on a microplate reader; after color development, measure within 15 minutes.
16. Calculate the sample content according to the prepared standard curve human ELISA kit
4. Results judgment
1. Instrument value: read the OD value of each well on a microplate reader with a wavelength of 450nm;
2. Detection value range: 0-400pmol / L
3. Sensitivity: 1.0pmol / L
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