Macro-Prep

Ion exchange chromatographies were carried out first on a Macro-Prep High Q (IBF) column (10 ml) in Tris-HCl 20 mM (pH 8), then on a 5 ml Econo-Pac Q cartridge (BioRad) equilibrated in ammonium carbonate 20 mM buffer (pH 5) and on a 5 ml Econo-Pac S cartridge (BioRad) equilibrated in ammonium carbonate 20 rnM buffer (pH 5). 

Protein extracts were eluted through HiTrap 1-mL columns, prepacked with one of four HIC resins methyl support (Macro-Prep HIC supports Bio-Rad, Hercules, CA) HiTrap HIC fast flow (FF) columns (Amersham Biosciences, Piscataway, NJ) butyl 4 FF octyl 4 FF phenyl 6 FF (low sub) sepharose. From every pelleted cell culture, 250-pL aliquots TPP extract were mixed with 250 pL of 4 M (NH4)2S04 and transferred to the top of each of the four FF support columns. The columns were previously... 

Macro-Prep DEAE Hydrophilic polyether ca. 50 Diethylaminoethyl 4-8 Weak anion-exchanger... 

Macro-Prep Ceramic Hydroxylapatite (Ca3(P04)2l3Ca(0H)2 20,40,80 Ca2+, P02 5.5-10 Weak cation- and anion-exchanger... 

Apply the sample at 2 ml/min to a hydroxyapatite (Bio-Rad Macro-Prep Ceramic hydroxyapatite, Type I) HR 10/10 column (Pharmacia) equilibrated with buffer B. Wash the column with 80 ml of buffer B develop the column with a linear gradient from buffer B to buffer B - - 1 M K2HPO4/KH2PO4 pH 7.5 over 80 ml at a flow rate of 2 ml/min. saTBP elutes as the major peak centered at 440 mM K2HPO4/KH2PO4. 

Macro-Prep High Q PolyacrylamirJe 50 Quaternary aminomethyl... 

Macro-Prep methyl divinylbenzene Methacrylate 50 1-14 n.a. Methyl >25 HSA molecular-weight molecules Bio-Rad... 

Macro-Prep Ceramic Calcium phosphate 20 40 80 Hydroxyapatite Type I... 

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