[Histonet] HPLC and Protein extraction
Have just embarked upon HPLC analysis for 5-HT and 5-HIAA in lizards.
Got some HPLC data that looked quite good (5-HT and 5-HIAA levels in fmol
being different in pharmacologically manipulated animals). But when i
normalized the data to amount of protein in the pellet, my results vanished
due to the differences in protein levels in my samples (which were all over
1.I figure that if i take the same number of punches per region per animal,
my protein levels should not vary drastically within a region. Is that a
wise assumption to make?
2. While performing the HPLC, i suspend my tissue punches in perchloric
acid and spin it down. Does anyone know of a protocol i might look into to
extract protein from this pellet. I'm presently using Sigma's Mammalian
Cell Lysis kit and then using a protein assay by Pierce Biotech?
Any input will be appreciated.
Brian George Dias
Graduate Student (Crews lab) Office Phone #: 512-475-6738
The University of Texas at
Austin E-mail: firstname.lastname@example.org
Institute for Neuroscience Website:
1 University Station Stop C0930
Austin, TX 78712
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