Cell Function
Cell Function FAQs
In order to provide better customer support and answer customers' questions about cell function assay products, we have compiled and published answers to the frequently-asked questions (FAQs) about cell function assay products, and will continue to update them.
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Heavy metal ions such as silver ions, copper ions, halogen ions and oxidizing organic compounds (nitro compounds, diazo compounds, carbonyl compounds and hydroxyl compounds) will quench the fluorescence of the fluorescent probe, thus affecting the accuracy of the experimental results.
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100 assays is calculated according to 96-well plate with 100μL working liquid per well, 24-well plate with 200μL working liquid per well, about 50 wells can be test.
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Matrix glue should be removed, not removed may interfere with calcein and cell binding; However, our calcein kit has not been verified by organoids, it is not clear what the final effect is, it is recommended to do a pre-experiment to see the actual effect.
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It's theoretically possible, but we haven't tested it, it is recommended to do a preliminary experiment to explore the details of the experiment.
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The cells of double positive are living cells in a poor state because the increased permeability of the cell membrane causes the nuclear dye to enter the cell. The cells of double negative are large granular cell fragments.
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The protein concentration was 1-4 mg/mL, and the caspase activity ranged from 2 U/ng prot to 430 U/ng prot.
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After the tissue sample is dry frozen, basically the cell morphology may have been destroyed and can not be used for flow detection. You can look at ELISA or metabolic indicators related to research, such as apoptosis of Bax, Caspase or oxidative stress of SOD, MDA and so on.
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① The microscope parameters are consistent; ② Dissolve the buffer and observe whether there are small red particles. If the buffer does not dissolve, centrifuge at 12000rpm and take the supernatant. (3) Apoptotic cells stick weakly to the flask and will float in the medium. If fluorescence microscopy is used, the results are relatively inaccurate, and flow cytometry is recommended for detection. Before collecting cells, take a photo at 100x magnification under normal white light (200x for small cells, such as lymphocytes) to check morphological changes.
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We do not recommend using tissue samples to prepare single-cell suspension for JC-1, because the operation of preparing single-cell suspension has a great impact on cells and will directly affect the experimental results of JC-1.
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It's theoretically possible, but we haven't tested it yet. We cannot determine the amount of JC-1 and the amount of extracted mitochondria, and may need to explore optimization.
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The monomers and polymers of JC-1 exist in both normal and apoptotic cells. In the process of cell apoptosis, JC-1 slowly dissociates from polymer to monomer, and the ratio of polymer to monomer changes. Therefore, when drawing the door, it is generally gathered in the third quadrant, and when drawing the door to distinguish between normal cells and apoptotic cells, it needs to be divided in the third quadrant.
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Generally, the absorbance value of CCK-8 is recommended to be about 1, preferably 0.8 to 1.2.
