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Lysis buffer for leptin receptor and pSTAT detection


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#1 Maria Boije

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Posted 25 November 2013 - 05:57 AM

Hi

 

I want to detect leptin receptor and pSTAT in tissue samples using western blot and would like some suggestions on lysis buffer and protein preparation procedure. I tried a buffer (20 mM Tris, 150 mM NaCl, 1 mM Na2EDTA, 1 mM EGTA, 1% Triton X-100, 2.5 mM Na4P2O7 with phosphatase and protease inhibitors) generally used in the lab for WB, but could only see a faint band at the estimated size of leptin receptor. In many papers I looked at where people have detected leptin receptor on WB they used RIPA buffer. Would RIPA buffer be OK for pSTAT detection? I am considering making first a preparation of cytosolic proteins and then one for membrane proteins. What buffers are best in that case? 

 

Thanks

Maria B



#2 bob1

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Posted 25 November 2013 - 12:02 PM

RIPA is a general lysis buffer that is fine for most proteins.  I don't know specifically for pStat though.






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