Homemade Luciferase Assay
1X Lysis Buffer
Add 1mM DTT just before use
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Substrate BufferFrom 1X lysis buffer to which add the following just before use:
Not essential: to boost the signal 2-3 fold you can also add 0.1 mM Coenzyme A. However, the background signal increases also accordingly.
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For cells grown in normal 96 well plates:
- Discard medium and tap the plate with paper towel to remove residual medium
- Add 100µl/well of 1X lysis buffer and incubate the plate on a shaker for 20 minutes at RT.
- Transfer 50µl of lysate to white plate.
- Add 50µl substrate buffer and read luminescence.
NB: if you rinse 4-5 times the white plates with tap water and once with distilled water and dry them, you can reuse the same plates indefinitely.