Protein sample is diluted with an equal volume of sample buffer. The mixture a boiling-water bath for 5 minutes. The amount of protein to be applied depends on the sample but applications in the range 1-5ug of protein per protein band usually give sharp easily viewed bands. Sample volumes of 10-100ul are most common Capacity per Well (Mini-Gel System) GelThickness 1Well 5 Wells 10 Wells 15 Wells 0.5mm 0.7ml 075mm 1.0m guI 4ul 1. 4ml 山 32μ!l 1.5mm 48μl Introduce sample solution into well using a Hamilton syringe(Fig 3). Layer protein solution on bottom of well and raise syringe tip as dye level rises. Be careful to avoid introducing air bubbles as this may allow some of sample to be carried to adjacent well. *Rinse syringe thoroughly with electrode buffer or water before loading different samples. Include molecular weight standards in one or both outside wells. Fig 3. Introducing protein solution into sample well 2. Molecular Weight standards: a slab gel is especially useful for molecular weight determinations since the sample and molecular weight standard proteins can be run under identical conditions on a single gel200 Protein sample is diluted with an equal volume of sample buffer. The mixture is heated in a boiling-water bath for 5 minutes. The amount of protein to be applied depends on the sample but applications in the range 1~5g of protein per protein band usually give sharp, easily viewed bands. Sample volumes of 10~100l are most common. *Capacity per Well (Mini-Gel System) Gel Thickness 1 Well 5 Wells 10 Wells 15 Wells 0.5mm 0.7ml 45l 16l 9l 0.75mm 1.0ml 68l 24l 14l 1.0mm 1.4ml 90l 32l 18l 1.5mm 2.1ml 135l 48l 27l *Introduce sample solution into well using a Hamilton syringe (Fig 3). Layer protein solution on bottom of well and raise syringe tip as dye level rises. Be careful to avoid introducing air bubbles as this may allow some of sample to be carried to adjacent well. *Rinse syringe thoroughly with electrode buffer or water before loading different samples. Include molecular weight standards in one or both outside wells. Fig 3. Introducing protein solution into sample well. 2. Molecular Weight Standards: A slab gel is especially useful for molecular weight determinations since the sample and molecular weight standard proteins can be run under identical conditions on a single gel