Like One gram of shoot and root tissue need to homogenize in 4 ml of 100 mM potassium phosphate buffer (pH 7.4) containing 1 mM ethylenediaminetetraacetate (EDTA), 2 % polyvinylpyrrolidone, and 1 mM phenylmethylsulfonyl fluoride (PMSF).
My question is how will I prepare this reaction mixture? Will I dissolve all [1 mM ethylenediaminetetraacetate (EDTA), 2 % polyvinylpyrrolidone, and 1 mM phenylmethylsulfonyl fluoride (PMSF)] in 100mM potassium phosphate buffer (pH 7.4). or I need to prepare all of this separately.
And in case of SOD
will for the reaction mixture (2.5 ml) : 75 nm riboflavin, 2 mM NBT, 3 mM EDTA, 200 mM methionine need to dissolve in 0.1 M potassium phosphate buffer (pH 7.8)? or I need to prepare all of this separately.
Please suggest me and help me out of this situation, as I never did this experiment before.
Thank you
The best is to work with stock solutions so you can mix these into water for the desired composition of the extraction buffer and reaction mixture. The phosphate buffer can be 1M or 0.5M as stock solution, most other compounds 100x stock. PVP you can weigh in as powder.
plant enzymes are stimulated or inhibited depending on climate change or pollutants
SOD/XOD is better enzyme to study the stress of the plant but my only dought is its stability during extraction of enzymes . Water hyacinth it is easy to study in the lab by adding different inorganic salts Cr, Fe, Cd etc
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