The animal serum manufacturer tells you the purpose of the sample
Animal serum: place the whole blood sample collected in the serum separation tube at room temperature for 5-2 hours or overnight at 4°C, then centrifuge at 1000×g for 20 minutes, take the supernatant, or remove the supernatant at -20°C or -80 ℃ but avoid repeated freezing and thawing.
Plasma: Use EDTA or heparin as an anticoagulant to collect specimens, centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes after collection, take the supernatant for testing, or store the supernatant at -20°C or -80°C but avoid Freeze and thaw repeatedly.
Tissue homogenate: Rinse the tissue with pre-cooled PBS (0.01M, pH=7.4), remove residual blood (lysed red blood cells in the homogeneous slurry will affect the measurement results), and cut the tissue after weighing. Cut the tissue and the corresponding volume of PBS (generally according to the weight-to-volume ratio of 1:9, such as 1g tissue sample corresponds to 9mL of PBS, the specific volume can be adjusted according to the needs of the experiment and recorded. Add protease inhibitors to PBS), Grind well on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly freeze-thawed. Finally, add the homogeneous slurry to 50,000 × g and centrifuge for 5-10 minutes, take the supernatant for testing, or store the supernatant at -20 minutes at -80°C but avoid repeated freezing and thawing.
Cell lysate: Discard the culture medium PBS (0.01M, pH7.4) and wash the cells again. Use cell scraper to remove cells (trypsin and trypsin cannot be used for EDTA treatment), add 2-5mL PBS (0.01M, pH7.4). Suspension cells can be omitted. Collect the cell suspension, centrifuge at 1000×g at 4°C for 10 min, discard the medium, and rinse with pre-cooled PBS for 3 times. Add an appropriate amount of pre-cooled PBS or non-denaturing cell lysate (add protease inhibitors before use) to resuspend the cells. Usually, the amount of cells in one well of a 6-well plate requires 150-250 μL of PBS to resuspend. Place the sample at -20°C or -80°C, freeze the sample, then thaw the sample at room temperature, and freeze the sample 3 times repeatedly to completely decompose the cells, or ultrasonically disrupt the sample to achieve the purpose of lysis. Centrifuge at 10,000×g for 10 min to remove cell debris, take the supernatant for detection, or store the supernatant at -20°C or -80°C but avoid repeated freezing and thawing.
For supernatant cell culture or other biological samples, please centrifuge at 10,000×g for 20 minutes, take the supernatant for testing, or store the supernatant at -20°C or -80°C but avoid repeated freezing and thawing.
Sample storage and stability:
Samples can be stored at 2-8°C for 72 hours, and at -20°C or -80°C for more than 6 months. After the sample is collected, it is not a one-time test. Please freeze it according to the amount used once to avoid repeated freezing.
