Tricine coomasie stain
first question for me here.
please take a look at my image.
i made a standard tris-tricine gel in order to see expression of two small mol wight proteins that i hope i sucssesfully transfected into L8 cells.
this was my first tricine gel.
i then fixed it using gluteraldehyde and then stained with coomasie blue. yes i know coomasie is not the most sensitive method, i just feared i might have been losing my 7 and 9 KD proteins somehow in the transfere.
anyway it seems as if all the proteins are gettign stuck before reaching the 20 kd marker.
i might have added too much protein having loaded about 60 micrograms.. but the upper sizes seemed to resolve nicely. so i dont think thats the problem.
i used according to instructions, both a cothod and anode buffer etc...
i ran them for about 2.5 h @ 50-60 mAmp.
please any ideas? i wanted to see the expression of my protein domains.
by the end the buffer was slightly warm, but not hot...
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Tricine gel problem
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