Dear Md. Nazmul Hasan,

We regularly perform reporter assays in our lab. We follow the protocol provided with Promega Luciferase Assay System (E1500) for reporter assay. The protocol is very simple. However, you may need to standardize the cell lysis method. For T47D and MCF-7 cell lines, I keep the cells in cell lysis buffer  for 10 min rocking condition followed by scraping. I also skip the following centrifugation step as I found it redundant after multiple trials. Also you might need to optimize the amount of lysis buffer to be added. For 80-90% confluent cells grown in 24 well plates, I give 60-80 ul of the lysis buffer.

Also take special care of the storage and preparation of the stock solutions. The LAR II reagent is highly light and heat-sensitive. It is stable for 1 year if stored in -80oC. So, plan your experiments in a such way, so that you can utilize the kit optimally for 1 year provided you strictly maintained the storage conditions. Also do not delay taking readings after you add LAR II in the lysates. Try to finish the experiment within 1-2 minutes after you add LAR II.

For any other queries, please feel free to ask.

Best of luck 

 Dear Pallabi Sengupta,

Thank you very much. It will help me a lot. In need, I will communicate with you in future.  Regards.