Rat mucin (MUC5AC) ELISA kit instruction manual
2021-05-20 13:03:52
Shanghai Xitang Biotechnology Co., Ltd. 021-55229872, 65333639
Rat mucin (MUC5AC) ELISA kit instruction manual
 ( used in serum, plasma, cell culture supernatants and other biological fluids )
principle
This experiment used double antibody sandwich ABC-ELISA. The anti-rat MUC5AC monoclonal antibody was coated on the plate, the MUC5AC in the standard and the sample was combined with the mAb, and biotinylated anti-rat MUC5AC was added to form an immune complex attached to the plate, horseradish peroxidation. The enzyme-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 MUC5AC concentration is directly proportional to the OD value, and the MUC5AC in the specimen can be obtained by drawing a standard curve. concentration.
Kit composition ( 2-8 ° C preservation)
Coated Wells | 96 holes | Enzyme Conjugate | 12ml |
10× specimen dilution (Sample Buffer) | 12ml | 20×Wash Buffer | 50ml |
Standards: 200ng/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, etc., as soon as possible, store at 2-8 ° C for 48 hours; longer time must be frozen (-20 ° C or -70 ° C) to avoid Repeated freezing and thawing.
2. Standard solution preparation: Add 0.5 ml of distilled water before use and mix to form a 400 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 400 ng/ml standard solution to the first tube, mix and aspirate 500 ul with a pipette, 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 to each well and let it react at 37 ° C for 15 minutes in the dark.
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. Using standard products 40, 20, 10, 5, 2.5, 1.25, 0.625, 0 ng/ml as the abscissa and OD as the ordinate, plot on the coordinate paper and draw the standard curve.
3. Find the corresponding MUC5AC content on the graph based on the sample OD value.
Kit performance
1. Sensitivity: The minimum MUC5AC detection concentration is less than 0.3 ng/ml.
2. Specificity: Recombinant or natural rat MUC5AC can be detected simultaneously. Does not cross-react with other cytokines in rats.
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 4oC refrigerator.
5. This kit is for scientific research only and cannot be used for clinical diagnosis!
This device is used to delivery supplemental oxygen to a patient or person in need of extra oxygen.
Nasal oxygen cannula is made from medical grade PVC, consists of connector (universal connector is available), oxygen tubing (usually the length is 2M), nose tip, Y-shape connecting tube.
Features:
1. Green color and white transparent color are both available;
2. Transparent clear tube is helpful for inspection of gas delivery;
3. DEHP Free PVC is available;
4. Softer nose tip can be chosen;
5. Available of three basic sizes, adult, pediatric and infant, and if needed, neonate size is available;
BPAP HF,Niv Mechanical Ventilation,Non Mechanical Ventilation,Non Invasive Pressure Ventilation
Shenyang RMS MEDICAL TECH CO.,LTD , https://www.medicalrms.com