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  #1  
Old 05-16-2012, 07:22 AM
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Question Sds page



hi i am working on SDS PAGE. i am loasing BSA as a sample. 1st time when i did it, i observed bands. i have used silver staining method. but after that i have tried brilliant blue staining method. i have not obtained bands anytime. i have used 6% and 4 % gel. both the times it was failed.


please give me the possible reasons for that.


yashvi
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Old 05-17-2012, 06:46 AM
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Default Re: Sds page

How much protein are you loading per well? and do you see your protein ladder?
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Old 05-18-2012, 05:20 AM
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Default Re: Sds page

1mg/ml

i have not loaded protein ladder
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Old 05-18-2012, 08:09 AM
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Default Re: Sds page

1mg/ml is only the concentration, what is the amount of protein you loaded? It is always rules of thumb to load protein ladder.

Please tell us detail brilliant blue used; G250? R250?

And also include your staining protocol would be great. To be on the safe side, do include what type of PAGE, percent and running protocol. Do include formula for your sample buffer.

For BSA, as far as I know normal standard staining procedure should be more than sufficient.

With the information which you will provide here, will help us to troubleshoot what is the exact problem. Cheers!
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Old 05-18-2012, 08:12 AM
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Default Re: Sds page

1mg/ml is only the concentration, what is the amount of protein you loaded? For example you loaded 10uL thus 1mg/ml*0.010ml=X mg. X mg is the number I am looking for. It is always rules of thumb to load protein ladder.

Do include formula for your sample buffer and running buffer too.
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