Tubulin: An Unequivocable Efficiency!

Samples were extracted by using a slightly modified version of the protocol of Pettersson and Kiessling (1984). A 12.5?mL-aliquot of distilled water was added to 2.5?g of finely ground Tubulin sample (through a 1?mm screen) and allowed to rehydrate for 1?h at room temperature. Then, we added 5?mL of 3?mol/L HCl and 40?mL of ethanol. Samples were mixed, boiled, and cooled. Extracts were filtered by gravity through folded filter paper. SepPak C-18 cartridges (Nihon Waters K. K., Tokyo, Japan, No. WAT043395) were first equilibrated with 5?mL of methanol and washed with 5?mL of distilled water. Then, 1?mL of filtrate was mixed with 3?mL of distilled water and loaded onto a cartridge. The cartridges were then washed with 2?mL of 20% methanol and eluted with 3?mL of 80% methanol. Next, the extract was made up to 3?mL with 80% methanol. Samples were stored at ?30��C until analysis. The quantitative analyses of phytoestrogens (daidzein, genistein, formononetin, coumestrol, biochanin-A) were determined by high-pressure liquid chromatrography (HPLC) analyses (Pettersson & Kiessling 1984) in triplicate. Differences Ivacaftor in the chemical composition among the diet materials and in the apparent digestibilities of DM, CP, SIP, DIP, ADIP, ether extract (EE), NDF, ADF, TDN and N balance among the diets were analyzed by one-way analysis of variance (ANOVA) and tested by the Tukey�CKramer multiple comparison test in SAS 9.2 (SAS Institute 2004), with significance level at P?

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