( gCostar Cat. No. 3903. Tel: 18004921110...)Luciferase Reporter Gene Cells and Recombinant Luciferase Dilutions Studied with the CLIPR Luciferase Assay Kit and the LMax Microplate Luminometer (MaxLine Application Note #38)

7. PCRE-luc-CHOK1 cell line generated by stable transfection of PCRE-luc plasmid (Stratagene, Catalog #219075) into CHOK1 cells (ATCC #CRL9618).

PREPARATION OF REAGENTS AND LUCIFERASE STANDARDS
The kit instructions were followed for preparation of the reagent.
1. PBS-BSA solution was prepared by adding 1% bovine serum albumin to Dulbeccos PBS 1X.

2. The firefly luciferase stock standard (1 mg/mL) was prepared in PBS-BSA. It was divided into 40 L aliquots and stored at -70C until needed.

3. For the dynamic range standard curve, the stock luciferase standard was diluted (10 L to 1000 L) in PBS-BSA, followed by 1-to-10 serial dilution in PBS-BSA. The diluted standards (10 g/mL to 10 pg/mL) were kept on ice until use.

4. For the low end standard curve, the stock luciferase standard was diluted (10 L to 1000 L) in PBS-BSA, followed by 2 additional 1-to100 serial dilutions. The final working standards were prepared by 1-to-50 dilution of the previous solution, followed by 1-to-2 serial dilutions. The diluted standards (0.625 to 20 pg/mL) were kept on ice until use.

5. The luciferase assay substrate (Component A) was reconstituted by adding 10 mL of luciferase assay buffer (Component B) to the vial, followed by gentle swirling and allowing it to stand at room temperature for approximately 1/2 hour. That substrate/buffer solution was then diluted 1:1 with PBS 1X. (Thus the final concentration was comparable to that intended in the kit instructions, where the reagent and cell suspension is mixed in a 1:1 ratio.)

PREPARATION OF THE LMAX
1. The M injector was cleaned by pumping 75% ethanol through the system and allowing it to stand for one hour, followed by rinsing generously with deionized water (5 washes, 99 cycles each).

2. The M injector was filled with the luciferase reagent by pumping 2 mL
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