Other biochemical reagent sample collection, processing and storage methods
1. Serum-Avoid any cell stimulation during the operation. Use test tubes free of pyrogens and endotoxins. After collecting blood, centrifuge at 1000 × g for 10 minutes to separate the serum and red blood cells quickly and carefully.
2. Plasma ----- EDTA, citrate and heparin plasma can be used for detection. Centrifuge at 1000 × g for 30 minutes to remove particles.
3. The cell supernatant --- 1000 × g centrifuged for 10 minutes to remove particles and polymers.
4. Tissue homogenate ----- Mash the tissue with appropriate amount of saline. Centrifuge at 1000 × g for 10 minutes, take the supernatant
5. Storage ------ If the sample is not used immediately, it should be divided into small parts and stored at -70 ℃ to avoid repeated freezing. If possible, do not use hemolysis or hyperlipidemia. If there are a large number of particles in the serum, centrifuge or filter before testing. Do not thaw at 37 ° C or higher. Thaw at room temperature and ensure that the sample is thawed evenly and adequately.
Preparation of other biochemical reagents1. Standards: Serial dilutions of standards should be prepared during the experiment and cannot be stored. Before dilution, the standard was mixed by shaking. The dilution ratio is as follows:
80 ng / ml
(No. 6 standard product)
The original concentration is directly added to 50ul without dilution.
40 ng / ml
(No. 5 standard product)
100ul of original standard is added to 100ul of standard dilution
20 ng / ml
(Standard 4)
Add 100ul of Standard No. 5 to 100ul of Standard Diluent
10 ng / ml
(Standard No. 3)
Add 100ul of Standard No. 4 to 100ul of Standard No. 4
5.0 ng / ml
(Standard No. 2)
Add 100ul of Standard No. 3 to 100ul of Standard Diluent
2.5 ng / ml
(No. 1 standard product)
Add 100ul of Standard No. 2 to 100ul of Standard No. 2
0 ng / ml
(Blank control)
The original concentration is directly added to 50ul without dilution.
2. Dilution of washing buffer (50 ×): 50-fold dilution with distilled water.
Operation steps of other biochemical reagents1. Before use, mix all reagents thoroughly. Don't make the liquid generate a lot of foam, so as to avoid adding a large number of bubbles during sample addition, which will cause errors in sample addition.
2. The number of slats required is determined by the number of samples to be tested plus the number of standard products. It is recommended to make multiple holes for each standard and blank hole. Each sample is determined according to its own quantity, and those that can use multiple holes can be used as much as possible. The specimen was diluted 1: 1 with the specimen dilution solution and then added 50ul to the reaction well.
Add 50ul of the diluted standard to the reaction well and 50ul of the sample to be tested in the reaction well. Immediately add 50ul of biotin-labeled antibody. Cover the membrane plate, gently shake and mix, and incubate at 37 ° C for 1 hour.boys backpack
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