1. 1 Expression Standard Ken Conover March 2006

2. 2 Expression Standard Protein Purification SDS-PAGE Protocol His-Tag & Gel Code Blue Stains Making the Expression Standard Software Demo

3. 3 Protein Purification SDS-PAGE Protocol Prepare SDS-PAGE Apparatus Protein Sample Preparation Run SDS-PAGE Gel Stain and Destain Gel

4. 4 Which Stain to Use? His-Tag Stain –Binds the his-tag portion of the protein –Is moles vs. molecular weight an issue? Gel Code Blue Stain –Binds the entire protein –Binds all proteins

5. 5 µ grams Vs. moles Gel Code Blue Stain His-Tag Stain 0.5 1.0 2.0 µg x/3 2x/3 x x = 1.86e-10 moles x x

6. 6 E Standard Protocol Determine concentration of each protein used in the standard. Using the dilution formula (M 1 V 1 = M 2 V 2 ), vary protein concentration within the range (µg) of the expression standard. V1

7. 7 Thank You Rong Haleema Kellie