There are a number of commercially available SDS-PAGE molecular weight standards hich give a good spread of molecular weight lines in a gel. Molecular weight standards Mid-Range protein Protein Molecular Weight(kD) Phosphorylase b(rabbit Muscle) 97,4 BSA(Bovine Serum Albumin) Ovalbumin(Chicken Egg) Carbonic anhydrase 31 Lysozyme( chicken Egg white 14.4 IV SLAB ELECTROPHORESIS CONDITIONS Running a gel 1. Attach electrode plugs to electrodes. Current should flow towards the anode 2. Turn on power supply to constant voltage; amperage will be about 100mA at start, 60mA at end of electropl for two 0.75mm gels: 110mA at start. 80mA at end for two 1. 5mm gels) 3. The dye front should migrate to lcm from the bottom of the gel in 30-40 min for two 0. 75mm gels(40-50 min for 1. 5mm gels) The high electrical current used in gel electrophoresis is very dangerous. Never disconnect electrodes before first turning off the power source If using an electrophoresis apparatus which is not completely shielded from the environment, always leave a clearly visible sign warning that electrophoresis is in progress. 4. Turn off power supply 5. Remove electrode plugs from electrodes. 6. Remove gel plates from electrode assembly. 7. Carefully remove a spacer, and inserting the spacer in one corner between the plates, gently pry apart the gel plates. The gel will stick to one of the plates.201 There are a number of commercially available SDS-PAGE molecular weight standards which give a good spread of molecular weight lines in a gel. *Molecular Weight Standards Mid-Range Protein: Protein Molecular Weight(kD) Phosphorylase b(Rabbit Muscle) 97.4 BSA(Bovine Serum Albumin) 66 Ovalbumin(Chicken Egg) 43 Carbonic anhydrase (Bovine Erythrocytes) 31 Lysozyme(Chicken Egg White) 14.4 IV. SLAB ELECTROPHORESIS CONDITIONS: Running a Gel *Steps 1. Attach electrode plugs to proper electrodes. Current should flow towards the anode. 2. Turn on power supply to 200V (constant voltage; amperage will be about 100mA at start, 60mA at end of electrophoresis for two 0.75mm gels; 110mA at start, 80mA at end for two 1.5mm gels). 3. The dye front should migrate to 1cm from the bottom of the gel in 30-40 min for two 0.75mm gels (40-50 min for 1.5mm gels). *The high electrical current used in gel electrophoresis is very dangerous. Never disconnect electrodes before first turning off the power source. If using an electrophoresis apparatus which is not completely shielded from the environment, always leave a clearly visible sign warning that electrophoresis is in progress. 4. Turn off power supply. 5. Remove electrode plugs from electrodes. 6. Remove gel plates from electrode assembly. 7. Carefully remove a spacer, and inserting the spacer in one corner between the plates, gently pry apart the gel plates. The gel will stick to one of the plates