insulin A on sds page? how! please help - (Mar/15/2007 )

hi all,

thanks to all who can solve my problem
i want to run insulin A on sds page for my experiment
but to till now i was not able to get a band for insulin A
i was able to to get a band for insulin B .
i even tried on tricine sds page but had no use
if there is any other way to resove it
can anyone please give a proper suggestion for the same???
i will be very greatful ...
waiting for a quick reply


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What percent tricine gel are you running? Try increasing the percentage, or increase the ratio of bis-acrylamide in the mix. This will decrease the pore size, which should allow better retention and resolution.

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thanks for the reply
i have already run it on 16.5 % gel
with4%T 3%C for stacking
10%T3%C for spacer
16.5%T 6%c for resolving

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Try to prepare Acrilamide stock solution 30%acrylamide /0.8% bisacrylamide ( commonly used for analytical gels 30/2.7) to improve resolution of your PAAGs

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30.8% will not work out .. i am running insulin !!!!!!

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Hi! Nish!

First of all not 30,8% but 30% acrylamide +0,8% bis acrylamide -- as a stock solution. To resolve insulin chains run Tricine PAAG ( 20%) or better 10-20% gradient

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ok u mean to say that i use 20% gel and try out the resolution.
one more thing about conjugation of cBSA and estradiol.. how can i confirm the conjugation

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I mean 20% or 10-20% gradient TRICINE PAGE

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ok i will try that

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