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Q：What is the difference in the number of suspended cells and adherent cells?
Because of the difficulty of staining suspension cells, it is necessary to increase the number of cells and extend the time of culture. Staining of adherent cells is relatively easy, and if the number of cells is too large, sometimes the absorbance will exceed the reading of the microplate reader.
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Q：How many cells should be seeded in each well for a live cell test?
When standard 96-well plates were used, the minimum inoculum size of adherent cells was at least 1,000 cells per well (100 μl of medium) . The sensitivity for detecting white blood cells is relatively low, so a dose of no less than 2,500 cells per well (100 μl of medium) is recommended.
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Q：Can Cck-8 stain living cells?
No. Because the major component of CCK-8 is a water-soluble tetrazolium salt (WST-8) , and electrons in living cells are exchanged by the electron carrier PMS onto WST8 in the medium. CCK-8 can not stain cells because the resulting jazzans are also highly water-soluble.
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Q：How can the error due to CCK-8 residue on the gun head or hole wall be reduced?
CCK-8 reagent can be diluted with medium and mixed before adding sample.
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Q：Is it necessary to test whether the medium and CCK-8 react before the experiment?
If necessary, check it with a hole. Since there may be a redox substance in the medium, it is necessary to confirm that the medium reacts with CCK-8 before a formal experiment. The normal OD should be below 0.4.
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Q：CAN CCK-8 detect bacterial cells?
You can test for E. E. coli, but could not detect yeast cells. To 100 ΜL. Coli culture medium was added with 10 μl CCK-8 solution and cultured for 1-4 hours or overnight.
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Q：Storage conditions for CCK-8 reagent?
The storage condition of CCK-8 kit of Liji Biology: 4 ° C short-term storage, valid for 3 months; -20 ° C long-term storage, valid for 24 months. [ note ] : repeated freezing and thawing will result in higher background OD values.
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Q：CCK-8 test solution toxic to cells?
The CCK-8 solution itself is slightly toxic due to the presence of high concentrations of PMS. However, CCK-8 added to the medium was not toxic because it was diluted 10-fold. Therefore, prolonged incubation, such as overnight or for several days, is acceptable. The same cell culture medium after CCK -8 detection can also be used for other cell proliferation detection, such as crystal violet detection, neutral red detection or DNA fluorescence detection. Since each cell has a different tolerance to CCK-8, the viability of the cells after addition of CCK-8 in culture is first examined when a long period of culture is required.
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Q：Is there a difference between skim milk and BSA seal?
In general, BSA produces clearer results because it has fewer proteins and antibodies have less chance of cross-reacting with it. However, this is not always the case. Some antibodies work better in skim milk, which contains more types of blocking proteins and can block more different types of proteins.
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Q：How many cells do WB experiments typically require?
5 × 106 is usually sufficient. But depending on which cell it is, some cells are small and some are spread out. It takes 105 cells to perform routine WB on a single pore. But it is also related to the target protein and the method of extraction. If it is a membrane protein or an organelle distribution of protein cells have increased.
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