Amount of 2-mercaptoethanol in lysis buffer for western blot

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yyua042
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Amount of 2-mercaptoethanol in lysis buffer for western blot

Post by yyua042 » Thu Jul 12, 2012 3:17 am

Hi all.

I added 20uL of 2-mercaptoethanol in 600uL of lysis buffer (3% volume), and I used this to do my western and I got no signals on my membrane.

I dont know if the reason why I got no signals is because that 3% of 2-mercaptoethanol was too low.

How much do you guys add into your lysis buffer?

Thanks

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JackBean
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Post by JackBean » Thu Jul 12, 2012 7:48 pm

Why should this matter?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

yyua042
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Re: Amount of 2-mercaptoethanol in lysis buffer for western blot

Post by yyua042 » Thu Jul 12, 2012 9:40 pm

My sample became sticky even after boiling for 10mins at 95degree and I wonder if this is because of the low volume of 2ME in the sample

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JackBean
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Post by JackBean » Fri Jul 13, 2012 7:21 am

Sticky? I would expact rather lack of water than of bME :lol:
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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