container. itil solution reaches top of front pl 5. Carefully insert comb into gel sandwich until bottom of teeth reach top of front plate (Fig 2). Be sure no bubbles are trapped on ends of teeth. Tilting the comb at a slight angle is helpful for insertion without trapping air bubbles. 6. Allow stacking gel to polymerize(about 30 minutes) 7. After stacking gel has polymerized, remove comb carefully(making sure not to tear the well ears) 8. Place gel into electrophoresis chamber 9. Add electrophoresis buffer to inner and outer reservoir, making sure that both top and bottom of gel are immersed in buffer. ig 2. Inserting the ab into the stacking gel. IIL PROTEIN SAMPLE PREPARATION 2.5ml Mercaptoethanol, 10ml Upper buffe Added H20 to 50ml199 container. 4. Pipet stacking gel solution onto separating gel until solution reaches top of front plate. 5. Carefully insert comb into gel sandwich until bottom of teeth reach top of front plate (Fig 2). Be sure no bubbles are trapped on ends of teeth. Tilting the comb at a slight angle is helpful for insertion without trapping air bubbles. 6. Allow stacking gel to polymerize (about 30 minutes). 7. After stacking gel has polymerized, remove comb carefully (making sure not to tear the well ears). 8. Place gel into electrophoresis chamber. 9. Add electrophoresis buffer to inner and outer reservoir, making sure that both top and bottom of gel are immersed in buffer. Fig 2. Inserting the sample-well comb into the stacking gel. III.PROTEIN SAMPLE PREPARATION: 1. Sample buffer: 1g SDS, 5ml Glycerol, 0.5mg Bromophenol blue, 2.5ml Mercaptoethanol, 10ml Upper buffer, Added H2O to 50ml