Synonym(s): RBC Lysing Buffer
Application: 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.
Components: Contains 8.3 g/L ammonium chloride in 0.01 M Tris-HCl buffer.
Other Notes: Note: This product is intended for the removal of red blood cells from mice. This product may not be appropriate for the lysis of red blood cells of other animals. The suitability of the product in any application other than mouse splenocytes must be determined by the researcher.
Legal Information: Hybri-Max is a trademark of Sigma-Aldrich Co. LLC
Biochem/physiol Actions: 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.
Empirical Formula (Hill Notation): C3H4Cl2O