I would like extract the oxidative enzymes from plant, but I cannot use the polyvinylpyrrolidone (PVP) for extraction. Is there another protocol, where I cant without PVP?
You can use polyvinylpolypyrolidone instead of polyvinylpyrrolidone. If the material from which you are extracting enzymes does not have negligible level or no phenolics, it is not necessary to have PVP or PVPP. Can you let me know the reason, why you do not wish to use PVP? PVPP has very good potential to adsorb phenolics.
thank you dear Pardha, but I have not this material (PVP) in my labor
Do you have lot of phenolics in your plant material? If phenolics are not a problem, you can use extraction buffer without PVP. If you need PVPP, I can donate some. You can send me your complete postal address. I shall be able to send, if there are no postal problems. Orelse you can make arrangements to pick it from my lab.
Without PVP, you will get hindrance of phenolics. You can try Tris-HCl Buffer (pH 7.5) for extracting the enzymes. Add PMSF (Phenyl methyl sulphonyl fluoride) to the extracting buffer just before the use. Perform the extraction at 4 degree Celsius in a cold room. You can also follow the protocol for extraction of individual enzymes as described by Mishra et al (Indian Journal of Biochemistry and Biophysics, 50 (2) (2013) 139-149).
You can see these articles (https://www.researchgate.net/publication/235989077_Copper-induced_changes_in_antioxidative_response_and_soluble_protein_level_in_Triticum_aestivum_cv._Beti_seedlings?ev=prf_pub: or http://www.gbm.bio.uaic.ro/index.php/gbm/article/viewFile/935/900).
We use some phosphate buffer for extraction.
Good luck!
Article Copper-induced changes in antioxidative response and soluble...
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