galactosidase assay for cells transfected with pSV- -Galactosidase Vector

Chen-Ming Fan Lab,Carnegie Institute of Washington

Prepare cell lysate as you would for Luciferase assay.

Take 50uL of lysate and dilute with 100uL of 1X

Reporter Lysis buffer (brown bottle at room temp cat#E397A) giving a total volume of 150uL.

Add 150uL of 2X assay buffer to each tube.

Vortex briefly.

Incubate at 37oC for 30 min or until yellow color has developed.

Stop the reaction by adding 500uL of 1M sodium Carbonate.

Your total volume is now 1mL.

Vortex

Read absorbance at 420 nm.

All components for b-gal assay are on top shelf of -20℃.

All components for Luciferase assay are in Common Rack B.

For info about this kit and map of pSV-b-gal vector, look in the cardboard box of kit manuals by Fan’s office.