Luciferase Assay!

Steve Crampton via methods%40net.bio.net (by scrampto from uci.edu)
Mon May 12 14:41:28 EST 2008


For luciferase assays, we always use the one-to-one proportion (100uL  
of lysate:100uL of substrate).  Looks like you were on the right  
track!  We also assay cells from 6-well plates and Steve


Message: 2
Date: Thu, 8 May 2008 20:01:10 +0530
From: "Sudheendra Rao N R" <sudhee26 from gmail.com>
Subject: Luciferase Assay!
To: methods <Methods from magpie.bio.indiana.edu>,
	methods from magpie.bio.indiana.edu
Message-ID:
	<a1a1abbb0805080731o1841b171xd20d37076f51b153 from mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Hi all,
1. I am doing Luciferase assay to understand transcriptional  
regulation. I
was looking for proper reporter lysis buffers.
i found out one with KH2PO4 (7.8 pH) with 0.2% Triton X-100 (v/v)..is  
there
any better?
it takes time for us to order promega 5x reporter lysis buffer.

2. i am doing duplicate assays in 6 well plates..and collecting cells  
in pbs
and lysing them in 200uL buffer. however i am not sure what is the
"standard" lysate:luciferase substrate (roche) proportion.
i have tried 10uL:90uL, 40uL:60uL, 100:100uL combinations, later gives  
me
better RLUs
we use a stand-alone, tube-luminometer (sirius), and do not use  
injectors.

please let me know.
(i will post my questions on DLuciferase Assay also..soon :))

thanks in advance
sudheendra.




More information about the Methods mailing list