Recommended for use by adding 1 mL of buffer to a cell pellet (cell pellet = 1 spleen or 100-200 million cells). Gently mix for 1 minute. Dilute the buffer with 15-20 mL of medium or salt solution. Centrifuge at 250-500 × g for 7 minutes and decant the supernatant. Cells may be diluted and prepared for counting or fusion. If lysis is incomplete, steps 1-4 may be repeated.
Red Blood Cell Lysing Buffer has been developed for use in hybridoma protocols to remove red blood cells from mouse splenocyte suspensions before fusion. It is also useful in systems where it may be desirable to remove red blood cells from cell suspensions, such as whole blood.
