Instructions for use
Fresh whole blood sample:
1. Add one volume of fresh whole blood and three volumes of red blood cell lysate. Add 3mL of red blood cell lysate to 1mL of fresh whole blood, gently swirl or invert and mix well.
2. Place on ice for 15 minutes, gently vortex and mix twice during this time. After the red blood cells lyse, the solution should be clear and transparent.
3. Collect cells: Centrifuge 450g at 4 ℃ for 10 minutes to precipitate white blood cells, and carefully discard the supernatant.
4. Add twice the initial blood volume of red blood cell lysate to the white blood cell precipitate, gently vortex and fully resuspend the white blood cells. If the initial blood volume is 1mL, add 2mL of red blood cell lysate.
5. Centrifuge at 450 g at 4 ℃ for 10 minutes to precipitate white blood cells, carefully and thoroughly remove the supernatant.
6. Resuspension cells for subsequent experiments; If RNA is extracted, it is best to start using a solution prepared with DEPC water at this step.
Fresh tissue samples:
1. Fresh tissue is digested and dispersed into a tissue cell suspension by collagenase and protease (or tumor tissue digest K601003), and the supernatant is discarded by centrifugation. 1mL of red blood cell lysate is added to the precipitate, and gently swirled or inverted to mix well.
2. Collect cells: Centrifuge 350g at 4 ℃ for 5 minutes to precipitate tissue cells, and carefully discard the supernatant.
3. Resuspension cells using PBS or basic culture medium for subsequent experiments.
Precautions:
This lysate is a sterile product. Please pay attention to the sterile operation when separating cells for cell culture. For your safety and health, please wear laboratory clothes and disposable gloves for operation.