how to prepare protein sample, if I want to load exact amount on SDS-PAGE

sudheer sangeetham via methods%40net.bio.net (by sudheer.pbm07 from gmail.com)
Fri Dec 16 16:01:24 EST 2011


Hello Guys
I have a trivial query, I have a protein stock 1000 µg per ml. Now I would
like to load on each well 0.25, 0.5, 1, 2, 4, 8, 16 and 32 µgs.
For 0.25  µg, 0.25  µl from stock + 0.25 µl 2x sample buffer , total 0.5 µl
load on first lane. If I want to load higher amount like 16 µg and 32 µgs
more volume to be loaded but I could load maximum16 µl in each well. In
that case, should I use C1 V1 = C2 V2 formula, 1000 µg * ? = 16 µg *  8 µl
V1 =  0.128 micro liter from the stock, 7.8 ul milli q water, to that add 8
µl 2x sample buffer and load 16 ul to the well, Am I right doing things
here?????

Thank you in advance

-- 
Sudheer Babu.S
Ph.D student
Institute of Biochemistry
Biological Research Center
Szeged,Hungary.


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