Protease

Just an update : I was being a dumbass ...sorry I just confused myself but thanks for the clarification and your patience. I did perform a 2d electrophoresis on the brain sample but loaded too much sample (I didn't think that silver staining would be as sensitive as it turned out to be). So far so good thanks again!

It's like a 2% agarose gel where I have the agarose but I am unsure if I need 100mL of EDTA or TAE to make it (solvent). I have all those chemicals but in solid form(CHAPS, Tris,Urea) and I want a solvent for those chemicals I was thinking either ddh20 or EDTA. Yet in that paper I listed above they have a ready made SolB (liquid) but in my case of I have to make a solution from solids.

In my lab the 7 M urea and 2M thiorea, CHAPS are all solids what would be a good base to use as a final volume liquid. In the paper they use Solution B which is composed of 7 M urea ..etc but I just have the chemicals in solid form and I was thinking either ddH20 or EDTA. Any recommendations? Note :The second step after homogenization would be delipidation using ethanol/acetone

Hi, I am currently working in a lab for the summer and the professor I have been working with wants to perform a 2D gel electrophoresis on rat brains. As an undergraduate, he has asked me to look up a procedure to perform 2d on rat brains which will assist him later on in his experiments, while it will help me get a good grade for coop/recommedation. I did find a procedure to homogenize rat brains but I am unsure of the amounts to use. I plan to use 0.01g of rat brain but in the article it says : Homogenization was performed in (Sol B) was: 7 M urea, 2 M thiourea, 40 mM Tris, 3 mM tributylphosphine,2% CHAPS, 1% Pharmalytes 3.5–10. How does one figure out how much to use for 0.01g of rat brain? I read through the article and no where does it mention how much to use (ul wise) to get good results when doing a 2d gel. Also, if anyone has a procedure of their own could post it here. The article I am using is Proteomic analysis of rat brain tissue: Comparison of protocols for two-dimensional gel electrophoresis analysis based on different solubilizing agents. Any assistance would be helpful. Thank you for your time. Regards