1 Expression Standard Ken Conover March 2006. 2 Expression Standard Protein Purification SDS-PAGE Protocol His-Tag & Gel Code Blue Stains Making the Expression.

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Presentation transcript:

1 Expression Standard Ken Conover March 2006

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

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

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

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 Thank You Rong Haleema Kellie