On Sat, 11 Jan 2003, Esk wrote:
> bear in mind if you're using ECL etc detection, you'll also have a challenge
> getting the bands for the two antibodies to come out at the same intensity
> with the same secondary ab concentration.
>In our experience this isn't the big problem. B-actin is more prevalent
than SNAP-25 so the real problem begins at the gel loading stage. If you
have enough SNAP-25 for the blot there will be too much B-actin. If you
decrease the amount loaded to obtain a favourable amount of B-actin, there
won't be enough SNAP-25. The only solution that worked for us for our
proteins (A-actin and an anti-apoptotic protein) is to load 2 gels in
parallel with adjusted volumes of lysate.
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legatek at mcmaster.ca Kyle Legate legatek at hotmail.com
Tower of Tongues:Thursday PM:10:30-11:30 EDT:http://cfmu.mcmaster.ca
moon musick:ritual:IDM:experimental(electronica):minimalism:glitch
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