Department of Physics, Chemistry and Biology (IFM)

Total protein was extracted from various tissues including brain, liver, skeletal muscle and heart of Dm-dNK positive and wild-type mice. The protein was extracted using RIPA Buffer (50 mM Tris HCl pH 7.6, 150 mM NaCl, 1% N-P40, 10% sodium deoxycholate, 0.1% sodium dodecyl sulphate (SDS) and protease inhibitors). Western blot was performed using 4-12% precast Bis Tris gel (NuPAGE) and Amersham Hyband-P membrane (Invitrogen). The presence of Dm-dNK protein was detected using anti-histidine antibody targeted against His-tag of the protein (1:3000) (CalBiochem) and anti mouse IgG linked to horse radish peroxidase (HRP) (1:3000) (GE Health Care). ECL (GE Health Care) was used as a substrate for the HRP.