Dear fellow research friends, i was to do biochemical assays of rat aorta and trachea,
Like sod and gpx, buti have no idea how much sample i should take for these biochemical assays, if you have protocols for this paper feel free to share?
Please keep in mind i studying muscle contraction by using organ bath.
You need to homogenize your sample in a suitable homogenizing buffer using an aliquot that is not less than 4 times the weight of your sample (the smaller the weight of the organ the higher the aliquot of homogenizing buffer). After homogenizing you centrifuge and decant the supernatant and store in a freezer. The kit or the laboratory procedure you are using will determine how much of the supernatant you will use for each parameter (e.g SOD, GPX etc). Please note that before each assay you will need to allow the supernatant to thaw and then keep in ice while working.
dear Shahnawaz Ahmad Wani,
for your question, it is very good to take 1 mg of each sample and proceed to enzymes extraction.
best regards
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