Can we use phosphate buffer pH 8 for tissue homogenate formation for different parameters of MDA ,reduced gultathione ,Catalase , SOD?
Sure you can! But most physiological proteins would be denatured at this pH. Thus, I would not expect 'huge' (typical) results. What are you trying to prove?
I agree with Bruce Neagle. I prefer to use pH having the optimal activity for an interested compound. For example, SOD has pH 7.8.
We also use a Phosphate buffer 0.1 M at pH 7.8 to run our SOD assays.
You can uses for CAT: phosphate buffer pH 7; for MDA: %1,15 KCl; for GPx: Tris- EDTA pH 7.4; for SOD: phosphate buffer pH: 7.4. Good luck.
for most of the parameter, we are generally used pH 7 to 7.4 for better enzyme stability. if u prepare tissue homogenates then go for the 0.1 M pot. phosphate buffer.
Hello Manisha.
They don't always have the same ph.
According to the experimental protocol.
Because the main purpose of using PB is to provide a good medium and maintain proteins and enzyme activity.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2830880/#R36
Hi,
I also prefer to use pH having the optimal activity for an interested compound.
In general I agree with Bruce Neagle. In my work I used Na/K-PO4 buffer pH 7.5 with for CAT and SOD activity determination as well as MDA (as TBARS method).
I use phosphate (Na-Na) buffer, pH 7 for CAT, SOD, GR and DHAR. Ka-Na phosphate buffer, pH 7 for TBARS.
Reduced glutathione you mean GSH or gluthatione reductase (GR)? If GSH I use 1% of
sulfosalicylic acid.
It depend of the enzymatic activity you want to measure . You have to prepare a special buffer for every activity for that I recommend to you to check the document
Gagné, F. (2014). Biochemical Ecotoxicology: Principals and Methods .Elsevier . https://doi.org/10.1017/CBO9781107415324.004
best regards
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