Dear Friends,

Our protein is purified cytokine 18 - 19 kDa we use disontinuous SDS PAGE 15% resolving and 5% Stacking. can some one give me the recipe for good Sample loading buffer for reducing and Non-reducing conditions. also we used mercaptoethanol for reducing condition and heated teh sample for 100 C / 5 min. is heating of sample necessary.

thanking in anticipation

exec

Here is a simple recipe that I have always used successfully

found on this website and in the "redbook"

http://www.bbri.org/faculty/smith/SDSBuffer.html

Rusty

* 25 mL buffer A
* 20 mL glycerol
* 4 g SDS
* (2 mL bME) add to aliquots note this would be 2% bME, Janet uses 10%
* 1 mg BfB
* H2O to 100 mL

Buffer A = 4X TrisCl/SDS pH 6.8 0.5 M TrisCl and 0.4% SDS
To make 0.5 mL of 2X SB, Red Book version

* 450 uL of SB
* 50 uL bME