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. Author manuscript; available in PMC: 2017 Apr 14.
Published in final edited form as: J Neurosci Methods. 2017 Feb 13;280:64–76. doi: 10.1016/j.jneumeth.2017.02.002

Figure 5. Comparison of optimized lysis buffer protein yield in directly homogenized and precipitated samples.

Figure 5

(A) Direct homogenization of frontal cortex with optimized lysis buffer (Opt. Homog) resolves significantly more protein per milligram of tissue than RIPA buffer (RIPA Homog). (A′) Representative Coomassie stained gels normalized for starting tissue mass. An arrow depicts a high molecular weight protein band that is only present if tissue is homogenized in optimized lysis buffer, and not RIPA buffer. (B) Optimized lysis buffer solubilization of TRIzol-precipitated protein (Opt. Precip) yields ~70% of the protein from hippocampal samples than if optimized lysis buffer is used for direct homogenization (Opt. Homog). (B′) Representative Coomassie stained gels normalized for starting tissue mass. Importantly, both the homogenized and precipitated protein samples resolve the protein band absent from RIPA buffer-homogenized protein. (C) Protein yields from (A) and (B) were normalized to optimized lysis buffer homogenate (Opt. Homog) within the same animal. Protein yield due to solubilization (Opt. Precip) is equivalent to protein yield in RIPA buffer homogenate (RIPA homog).