By Robert C. Dickson, Michael Dean Mendenhall
Sporting at the excessive criteria of the much-acclaimed first variation, hugely skilled investigators have widely up-to-date the 1st version with a few of the new methods which have been reworking the sphere. incorporated during this new version are simply reproducible immunoassays, fluorescence-based assays, high-throughput equipment, protein amendment assays, lipid moment messenger assays, and chromatin immunoprecipitation suggestions. anywhere attainable, protocols for the assay of normal periods of sign transduction parts were pointed out to allow them to be tailored to the assay of any member of that category.
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Additional info for Signal Transduction Protocols 2nd Editoin (Methods in Molecular Biology Vol 284)
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10. Reswell dehydrated gel pieces with 20 lg/mL Trypsin by adding solution directly onto gel pieces in 3–5 lL aliquots, waiting 10 min between each addition. Continue until gel pieces are fully hydrated (typically approx 10 lL). 11. After gel pieces are hydrated, add 25 mM ammonium bicarbonate to cover the gel pieces (approx 20 lL). Incubate at room temperature for 20 min. 12. When fully swelled, add 25 mM ammonium bicarbonate until gel pieces are covered at the top by approx 1 mm of buffer. 13.
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