Human follistatin analog 3 (FSTL3) ELISA kit
 ( used in other biological fluids such as serum, plasma, cell culture supernatant, and saliva )
principle
This experiment used double antibody sandwich ABC-ELISA. The anti-human FSTL3 monoclonal antibody was coated on the plate, the FSTL3 in the standard and the sample was combined with the monoclonal antibody, and biotinylated anti-human FSTL3 was added to form an immune complex attached to the plate, horseradish peroxidase. The labeled Streptavidin is combined with biotin, and the substrate working solution is blue. Finally, the stop solution sulfuric acid is added, and the OD value is measured at 450 nm. The concentration of FSTL3 is directly proportional to the OD value, and the concentration of FSTL3 in the sample can be obtained by drawing a standard curve.
Kit composition ( 2-8 ° C preservation)
Coated Wells | 96 holes | Enzyme Conjugate | 12ml |
10× specimen dilution (Sample Buffer) | 12ml | 20×Wash Buffer | 50ml |
Standards: 80ng / bottle | 2 bottles | Substrate working fluid (TMB Solution) | 12ml |
Primary antibody working solution (Biotinylated Antibody) | 12ml | Stop Solution | 12ml |
Prepare reagents and collect blood samples
1. Collect specimens: serum, plasma (EDTA), cell culture supernatant, tissue homogenate, saliva, etc., as soon as possible, store at 2-8 ° C for 48 hours; freeze for a longer time (-20 °C or -70 °C) Save to avoid repeated freezing and thawing. Pre-measure the normal specimen with a dilution of the specimen at least 1 : 50 (take 20ul, add 980ul of the diluted solution, diluted 50 times). Saliva can be detected directly without dilution.
2. Standard solution preparation: Add 2 ml of distilled water before use and mix well to prepare a 40 ng/ml solution. Set the standard tube 8 tube, the first tube plus the standard dilution 900ul, the second to the eighth tube to add the sample dilution 500ul. Add 100 ul of the standard solution of 40 ng/ml to the first tube, mix and aspirate 500 ul with the sampler, and transfer to the second tube. Repeat the dilution as described above, and remove 500 ul from the seventh tube and discard it. The eighth tube is a blank control.
3. The 10× specimen dilution was diluted 1:10 with distilled water (example: 1 ml concentrated dilution + 9 ml distilled water).
4. Washing solution: diluted 1:20 with distilled water (example: 1 ml concentrated washing solution added to 19 ml of distilled water)
Test procedure
1. Loading: Add 100 ul of standard or sample to be tested in each well. Mix the reaction plate thoroughly and let it stand at 37 °C for 120 minutes.
2. Wash the plate: Wash the plate thoroughly with washing solution 4-6 times, and dry it on the filter paper.
3. Add 100 ul of the first antibody working solution to each well. The reaction plate was thoroughly mixed and placed at 37 ° C for 60 minutes.
4. Wash the board: the SAMe as before. 5. Add 100 ul of enzyme-labeled antibody working solution per well. The reaction plate was placed at 37 ° C for 30 minutes.
6. Wash the board: same as before.
7. Add 100 ul of substrate working solution per well, set 37 The reaction was carried out in the dark at °C for 15 minutes.
8. Add 100 ul of stop solution to each well and mix.
9. Measure the absorbance at 450 nm using a microplate reader within 30 minutes.
Result calculation and judgment
1. All OD values ​​should be subtracted from the blank value before calculation.
2. Take the standard products 4000, 2000, 1000, 500, 250, 125, 62.5, 0 pg/ml as the abscissa and OD as the ordinate. Draw on the coordinate paper and draw the standard curve.
3. Find the corresponding FSTL3 content on the graph based on the OD value of the sample, and multiply by the dilution factor. .
Kit performance
1. Sensitivity: The minimum FSTL3 detection concentration is less than 30pg/ml.
2. Specificity: Recombinant or natural human FSTL3 can be detected simultaneously. Does not cross-react with other human cytokines.
3. Repeatability: The coefficient of variation in both the plate and the plate is less than 10%.
Precautions
1. It is recommended to make double holes for the above standard holes and samples to be tested. The standard curve should be made at the same time for each measurement.
2. The washing process is critical. Insufficient washing will result in an accuracy error and an erroneous rise in the OD value.
3. After the slats are opened, the remaining slats should be sealed again to keep the slats dry .
4. This kit should be stored in a 4 o C refrigerator.
5. This kit is for scientific research only and cannot be used for clinical diagnosis!
NMN For Capsule
When people was young, the human body had a good DNA damage repair mechanism, and can maintain a good cell renewal speed, in order to maintain the normal growth, development and reproduction of the body. With the increase of age, the level of DNA damage repair in human body decreased.
In addition, individual and environmental factors, such as stress and strain, injury and infection, decline of immune response, malnutrition, metabolic disorders, drug abuse, bad living habits, diseases, environmental pollution and so on, will accelerate the speed of DNA damage and reduce the level of repair.
NMN is a serious scientific research achievement that can significantly reverse aging and prolong life through rigorous scientific verification. With the deepening of research, more and more effects of NMN have been found, such as improving muscle aging, reversing hematopoietic stem cell activity, maintaining telomere length and so on.
Nmn For Capsule,99.5% Nmn Niacin,Nmn Powder 99%,Nmn Supplements
Yuyao Lifespan Health Technology Co., Ltd. , https://www.yuyaolifespan.com