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2 protocols using dna markers

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Purification and Detection of Human Prdx6 Protein

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Trizma Base, EDTA, Sodium Hydroxide, Sodium dodecyl sulphate, Ethidium bromide, Imidazole, Tween-20, Tween-80, Sodium chloride, Potassium chloride, Glycerol, Acrylamide, bis-acrylamide, Ammonium persulphate, TEMED, Glycine, β-mercaptoethanol, Hydrogen peroxide (H2O2), Dithiothreitol (DTT), Horseradish peroxidase (HRP), 5,5′-dithiobis(2-nitrobenzoate) (DTNB), Bovine carbonic anhydrase, and Alpha-lactalbumin were purchased from Sigma Aldrich Pvt. Ltd.
Luria Bertani Medium and Luria Bertani Agar (Difco), PBS tablets (Biobasic), Ni–NTA resin (Qiagen GmbH, Germany), DNA markers (GeneDireX Inc., USA), Precision dual colour Protein marker (Bio-Rad). Ampicillin and Kanamycin were purchased from MP Biomedicals.
Primary (Monoclonal Anti-Prdx6, clone 3A10-2A11 antibody produced in mouse, WH0009588M1) and secondary (Anti-mouse IgG (Fab Specific)-Peroxidase antibody produced in goat, A3682) to detect full length human Prdx6 protein was procured from Sigma-Aldrich.
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2

Purification and analysis of hPrdx6 protein

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Trizma Base, EDTA, sodium hydroxide, sodium dodecyl sulphate, glacial acetic acid, ethidium bromide, imidazole, sodium chloride, potassium chloride, glycerol, acrylamide, bis-acrylamide, ammonium persulphate, TEMED, glycine, bovine serum albumin, β-mercaptoethanol, hydrogen peroxide (H2O2), 8-anilino-1-naphthalenesulfonic acid ammonium salt (ANS), reduced glutathione (GSH), glutathione reductase, reduced β-nicotinamide adenine dinucleotide phosphate (β-NADPH), dithiothreitol (DTT), guanidinium chloride (GdmCl), luria bertani medium and luria bertani agar (BD Difco, USA), PBS tablets (Bio Basic), Ni-NTA resin (Qiagen GmbH, Germany), DNA markers (GeneDireX Inc., USA), precision dual colour protein marker (Bio-Rad), ampicillin and kanamycin (MP Biomedicals), HiYield PlusTM plasmid mini isolation kit (RBC Bioscience), bradford protein estimation kit (MP Biomedicals), EnzCheck phospholipase A2 assay kit (Invitrogen). Primary (monoclonal anti- hPrdx6, clone 3A10-2A11 antibody produced in mouse, WH0009588M1) and secondary (anti-mouse IgG (Fab Specific)-peroxidase antibody produced in goat, A3682) antibody procured from Sigma-Aldrich, E.coli strains: DH5α and M15[pREP4].
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