(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
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
<a1a1abbb0805080731o1841b171xd20d37076f51b153 from mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
1. I am doing Luciferase assay to understand transcriptional
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
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
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
we use a stand-alone, tube-luminometer (sirius), and do not use
please let me know.
(i will post my questions on DLuciferase Assay also..soon :))
thanks in advance
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