User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/06/03

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Tris urea gel
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Summary

 * Preparing 4x Urea loading buffer
 * 4x Wilfred´s Urea loading dye (10 mL)
 * 4.8g Urea (4.8 g)
 * 1.5224g Thiourea
 * 3.2mL stacking gel buffer (0.625M) Tris-HCl pH 6.8
 * 0.8g SDS
 * 0.3722g EDTA
 * 0.025g Xylene Cyanol
 * 0.025g Bromophenol Blue
 * 4mL glycerol
 * Up to 10 mL with ddH20


 * Glycerol might not be necessary due to the fact that the high Urea concentration will provide enough weight to the sample, SDS might also not be necessary.
 * Glycerol was left out due to volume limitation
 * SDS Did not seem to solubilize before adding the color reagents, afterwards this could not be checked
 * Before adding H2O to solubilize SDS I wanted to add the color reagents to be sure not to add to much volume
 * Sample RIPA lysate S0 used before was gone, so now sample from 15March2010 is used

Discussion

 * Perhaps the amount of glycerol prevented the sample to condense to a nice band, maybe we can try with normal running buffer so that glycerol does not need to be added


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