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Why does cell culture lyse red blood cells first?
2022-12-23
Basic introduction
Erythrocyte lysate is one of the most simple and easy methods to remove red blood cells, that is, to split red blood cells with lysate, which does not damage nucleated cells and can fully remove red blood cells. Lysate cleavage is a mild red blood cell removal method, which is mainly used for the separation and purification of tissue cells dispersed by enzyme digestion, the separation and purification of lymphocytes, and the removal of red blood cells in the experiments of tissue protein and nucleic acid extraction. The tissue cells obtained by the lysate of red blood cells do not contain red blood cells, and can be further used for primary culture, cell fusion, flow cytometry, separation and extraction of nucleic acid and protein, etc.
Instructions for use
Tissue cell sample
1. Fresh tissues were digested by pancreas/enzyme or collagenase and dispersed into single cell suspension, and the supernatant was discarded by centrifugation.
2. Take ELS lysate from the refrigerator at 4℃, add ELS lysate to cell precipitate in a ratio of 1:3-5 (add 3-5ml of lysate to 1ml of cell compacted), gently blow and mix.
3. Centrifuge at 800-1000rpm for 5-8 minutes and discard the upper red clear liquid.
4. The precipitated part was collected and centrifuged with Hank's solution or serum-free culture solution for 2-3 times.
5, if the cracking is not complete/complete can be repeated steps 2 and 3.
6. Resuspension cells for subsequent experiments; If RNA is extracted, it is best to do so in the solution prepared from Step 4 using DEPC water
Erythrocyte lysate is one of the most simple and easy methods to remove red blood cells, that is, to split red blood cells with lysate, which does not damage nucleated cells and can fully remove red blood cells. Lysate cleavage is a mild red blood cell removal method, which is mainly used for the separation and purification of tissue cells dispersed by enzyme digestion, the separation and purification of lymphocytes, and the removal of red blood cells in the experiments of tissue protein and nucleic acid extraction. The tissue cells obtained by the lysate of red blood cells do not contain red blood cells, and can be further used for primary culture, cell fusion, flow cytometry, separation and extraction of nucleic acid and protein, etc.
Instructions for use
Tissue cell sample
1. Fresh tissues were digested by pancreas/enzyme or collagenase and dispersed into single cell suspension, and the supernatant was discarded by centrifugation.
2. Take ELS lysate from the refrigerator at 4℃, add ELS lysate to cell precipitate in a ratio of 1:3-5 (add 3-5ml of lysate to 1ml of cell compacted), gently blow and mix.
3. Centrifuge at 800-1000rpm for 5-8 minutes and discard the upper red clear liquid.
4. The precipitated part was collected and centrifuged with Hank's solution or serum-free culture solution for 2-3 times.
5, if the cracking is not complete/complete can be repeated steps 2 and 3.
6. Resuspension cells for subsequent experiments; If RNA is extracted, it is best to do so in the solution prepared from Step 4 using DEPC water
Red blood cells have a very short life cycle, only 120 days, but they reproduce blood very quickly, and in this case they are particularly capable of cell division, and they are the fastest dividing cells of all, so this cell is very valuable, so it's very useful for cell culture. It's very simple, it doesn't have any organelles in it, just cell membranes and proteins.
