How can I make isolation buffer for cell isolation?
To prepare this buffer:
(1) Make up 100 mL of a stock solution of 0.5 M EDTA by dissolving 14.6 g EDTA in approximately 80 mL of ddH2O. Adjust the pH to 7.4, then adjust the volume to 100 mL. Autoclave and filter 25 mL aliquots through a sterilized filter into 4 sterile 50 mL tubes. Store one at 4 degrees C and the rest at -20 degrees C for future use.
(2) Make up a stock solution of 20% BSA by putting 5 g of BSA in a 50 mL tube and filling it with ddH2O up to 25 mL. Leave on a roller at room temperature until dissolved. Filter through a sterile 0.2 µm filter into a sterile 50 mL tube and adjust the volume to 30 mL with sterile H2O.
(3) Make the isolation buffer by taking a 500 mL bottle of 1x PBS, pH 7.4 without Ca2+ or Mg2+ (Cat. No. 10010023). Add 2.5 mL 20% BSA and 2 mL 0.5 M EDTA. Mix by turning the bottle end over end 10 times. Store at 4 degrees C.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
(1) Make up 100 mL of a stock solution of 0.5 M EDTA by dissolving 14.6 g EDTA in approximately 80 mL of ddH2O. Adjust the pH to 7.4, then adjust the volume to 100 mL. Autoclave and filter 25 mL aliquots through a sterilized filter into 4 sterile 50 mL tubes. Store one at 4 degrees C and the rest at -20 degrees C for future use.
(2) Make up a stock solution of 20% BSA by putting 5 g of BSA in a 50 mL tube and filling it with ddH2O up to 25 mL. Leave on a roller at room temperature until dissolved. Filter through a sterile 0.2 µm filter into a sterile 50 mL tube and adjust the volume to 30 mL with sterile H2O.
(3) Make the isolation buffer by taking a 500 mL bottle of 1x PBS, pH 7.4 without Ca2+ or Mg2+ (Cat. No. 10010023). Add 2.5 mL 20% BSA and 2 mL 0.5 M EDTA. Mix by turning the bottle end over end 10 times. Store at 4 degrees C.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.