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From
http://www.nationaldiagnostics.com/article_info.php/articles_id/6.
Retrieved 2/9/2012.
Polymerization of Acrylamide.
http://www.nationaldiagnostics.com/article_info.php/articles_id/6http://www.nationaldiagnostics.com/article_info.php/articles_id/6
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Typical Polyacrylamide Gel
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Protein Structure
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SDS Coats the Protein with a Uniform Negatie Charge
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Typical !esults
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Kaleidoscope and original prestained standards. Prestaining
proteinsalters their actual molecular weights, depending on the
dyes thatare used. Therefore, the apparent molecular weights will
varyslightly between the Kaleidoscope and original prestained
standards,as shown in this illustration.
Kaleidoscope re!"tained #olec$lar %eight #ar&ers
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%estern 'lot roced$re
1( )nc$*ate overnight at +,- in ' 10 dried mil&(
2( Rinse with '" a detergent to *rea& $p non!specific
*inding interactions(3 4 min
5( )nc$*ate at R.. in 2 ml mo$se anti!7$man "er$m 8l*$min
!50 min
+( Rinse *riefl with '"
( Rinse with 50 ml fresh '" 3 4 min 5
6( )nc$*ate with 2 ml anti!mo$se )g; 7R con
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http://www.*io!rad.com/en!$s/applications!
technologies/imm$nodetection> retrieved 9/20/2015
Detecting the Protein on the "lot' 10 dr mil&( ma&es
an effective? inefficient *loc&ing
agent. he mil& proteins *ind
the *lot so that the anti*odies
can onl *ind antigen? not the*lot.
http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=88KavKvL_Jg_LM&tbnid=w5nMu6a7eu7SdM:&ved=0CAUQjRw&url=http%3A%2F%2Fenvcal.blogspot.com%2F2013%2F04%2Fcarnation-instant-nonfat-dry-milk-44-lbs.html&ei=aY08UtOnLMfF2wWXnYDoCA&bvm=bv.52434380,d.aWc&psig=AFQjCNHBCKe5cSlQlnh9rcSGy2Wwi3o06Q&ust=1379786458661907
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#isualization of Anti$ody "inding to Target Protein
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BCHS3201: Serum Albumin Spring2009
Primary Antibody: Mouse Anti-humanalbumin; Secondary Antibody:
Goat Anti-
mouse IgG
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