Western Blots

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From:Ronnie Houston <wee_rory@hotmail.com>
Content-Type:text/plain; format=flowed

Not strictly a Histo question, but one of our researchers is doing some 
Western Blots for us on skeletal muscle samples from suspected Beckers 
Muscular Dystrophy.
She is using biotinylated standards. However, the primary dystrophin 
antibody is not biotin-conjugated, so she is applying a biotinylated 
secondary anti-mouse Ig to both test samples and the standard. This is 
followed by an avidin-conjugated alkaline phosphatase and visualization with 
My question is, can you compare the results of the standard to the test when 
the biotinylated standard has been subjected with IgG-biotin? Does this 
extra amount of biotin affect the result?
My initial thoughts are that one cannot directly compare results because 
differently prepared samples have been run. Also there has been no blocking 
for endogenous biotin.
Thanks for any help you can offer.

Ronnie Houston
Cytochemistry & Molecular Pathology
Texas Scottish Rite Hospital for Children
2222 Welborn Street
Dallas, TX 75219
(214) 559 7744
(214) 559 7768 - fax

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