LUHMES cryopreservation media 70% Advanced DMEM media 10% FBS 20% DMSO 2.1.2 Biochemicals a) Protein extraction and bicinchoninic acid (BCA) protein assay Extra strong lysis (XSLB) 10mM Tris-HCL (pH 7.5) 0.5% (w/v) sodium dodecyl sulphate (SDS) 0.5% (w/v) sodium deoxycholate 1% (v/v) Triton X-100, 75mM sodium chloride 10mM ethylenediaminetetraacetic acid 2mM sodium orthovanadate 1.25mM sodium fluoride Protease inhibitor cocktail (PIC) for mammalian tissues (1 tablet/ 10ml of XSLB) (Roche, Mannheim, Germany) Phosphatase inhibitor (1 tablet used/ 10ml XSLB) tablets (PhosSTOP™) (Roche, Mannheim, Germany) Bichinoic (BCA) protein assay BCA Protein Assay Kit (ThermoFisher Scientific, Waltham, MA, USA) Bovine serum albumin (BSA) standard standard (Pierce Endogen, Rockford, IL, Thermo Scientific™Pierce™ Albumin USA) b) Sodium Dodecyl Sulphate Polyacrylamide gel electrophoresis (SDS-PAGE) 30% Acrylamide/Bis solution (Bio-Rad Laboratories Ltd., Hertfordshire, UK) (Cat. No- A3678) Ammonium Persulphate (APS)
LUHMES cryopreservation media 70% Advanced DMEM media 10% FBS 20% DMSO 2.1.2 Biochemicals a) Protein extraction and bicinchoninic acid (BCA) protein assay Extra strong lysis (XSLB) 10mM Tris-HCL (pH 7.5) 0.5% (w/v) sodium dodecyl sulphate (SDS) 0.5% (w/v) sodium deoxycholate 1% (v/v) Triton X-100, 75mM sodium chloride 10mM ethylenediaminetetraacetic acid 2mM sodium orthovanadate 1.25mM sodium fluoride Protease inhibitor cocktail (PIC) for mammalian tissues (1 tablet/ 10ml of XSLB) (Roche, Mannheim, Germany) Phosphatase inhibitor tablets (PhosSTOP™) (1 tablet used/ 10ml XSLB) (Roche, Mannheim, Germany) Bichinoic (BCA) protein assay BCA Protein Assay Kit (ThermoFisher Scientific, Waltham, MA, USA) Bovine serum albumin (BSA) standard Thermo Scientific™Pierce™ Albumin standard (Pierce Endogen, Rockford, IL, USA) b) Sodium Dodecyl Sulphate Polyacrylamide gel electrophoresis (SDS-PAGE) 30% Acrylamide/Bis solution (Bio-Rad Laboratories Ltd., Hertfordshire, UK) Ammonium Persulphate (APS) (Cat. No- A3678)
FINAL%20thesis%20version%20jan%202017.docx