Lonza
ACK lysing buffer
10-548E
Info: Browse ACK Lysing Buffer Products
To study expression of different cell surface markers on immune cells during vaccination in mouse model, we prepare single-cell suspensions of mouse splenocytes, which contain a high population of red blood cells (RBCs). To have effective fluorochrome antibody binding, we need to lyse the RBCs present in splenocytes. To lyse the RBCs present in splenocytes we use ACK lysing buffer. This buffer lyses RBCs very effectively with no RBC contamination in mouse splenocytes cell suspension.
Red blood cells lysisng
Mouse splenocytes
Incubate loose pellet of mouse splenocytes single-cell suspension (around 100 to 150 million cells) in 4 mL ACK lysis buffer for 90 seconds. Add around 35 mL of FACS buffer or RPMI 1640 to neutralize the ACK lysis buffer. Centrifuge it in swinging bucket rotor for 5 mins, 4°C at 1500 rpm. Decant the supernatant. The cell pellet will be free from RBCs.
Add ACK lysis buffer after loosing the cell pellet.
Effectively lyse RBCs present in mouse splenocytes.
N/A
Can be used for RBCs lysis in human PBMCs but less effective
After lysing, splenocytes can be used for flow cytometry analysis, sorting, and cell culture.
Need more than one run to lyse blood RBCs
The ACK lysis buffer works effectively.