Heating with neutral phosphate buffer

Delignification of Spruce Wood Meal on Heating with Neutral Phosphate Buffer Solution... 

Tissues are fixed with 10% neutral phosphate-buffered formalin and embedded in paraffin, and sections (5 pm thick) are mounted on poly-L-lysine-coated slides heated at 60°C for 30 min. The sections are deparaffinized in four changes of xylene and then rehydrated in a descending series of ethanol. Endogenous peroxidase activity is quenched by immersing the sections in 1% H202 in distilled water for 5 min. The sections are rinsed in three changes of distilled water, transferred to a moist chamber, and covered with PBS. 

This quick assay is performed as follows (Verma et al., 1978) Retinoids and TPA are each dissolved in acetone and are applied separately to the shaved dorsal skin of mice in a volume of 0.2 ml. The mice are pretreated with the appropriate dose of retinoid 1 h before application of 17 nmol of TPA. The mice are sacrificed 4.5 h after dosing with TPA, which is the time of maximum ODC induction in the skin. The epidermis from individual mice is separated by brief heat treatment. The pooled epidermal preparations from three or four mice are homogenized in neutral phosphate buffer containing 0.1 mM pyridoxal phosphate and 0.1 n M EDTA and centrifuged briefly at high speed to obtain a clear extract. The ODC activity is determined on this extract by measuring the release... 

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