pplications Report: New Column Technology

Peak Identity Molecular Weight Column: Bro-SIl TSK-250 1 ThryrogJobuhn 670,000 300 χ 7 b mm 2. IgG 150.000 Flow Rater 1 ml nun 3. Ovalbumin 45 000 Buffer: 0 1 M N a S O . 4 Myoglobin 17,500 0Û2MNaH ? PO„ p H 6,8 5 Cyanocobalamin 1 3 5 5 Sample: Θιο-Rad Gel

Filtration Standard

Column" Βιο-Rad TSK-IËX-54D DEAE Sample: Β to-R ad Ciel Fil tration Standard

10μΙ inject ion Gradient:

Solvent A; 0.05M Trib Suffer, pH 7.2 Solvent B: 0 5M NaCI m 0.05 M Tris Buffer, pH 7 2 Linear gradient programmed from 0% to 100% Β for 45 minutes.

Flow Rate : 2 0 ml/mm Detector: UV at 280,0 nm Peak Identity:

1. IgG in solvent front 2 Myoglobin 3 Cyanocobalamm 4 Ovalbumin related peptide

S Ovalbumin related peptide 6. Ovalbumin , 7 Ovalbumin relatexJ peptide

Column: Β ιο-Rad Reverse Phase (RP-304) H)-Pore ,M

Sample: Β ιο-Rad Gel Filtration Standard, 10μΙ inject ion

Program: 1 100% A for four ο 25 -

minutes 2 Ltnear gradient from ω

0% to 100% Β over A minutes

Flow Rate: 0 6 mt>min Detector: υ ν at

2&0 0 nm Peak Identity;

1 Cyanocobaiamin 2 Ovalbumin

related impurity 3 4 Myoglobin 5 Protein impurity 6 Ovalbumin

Gradient: Solvent A: / 5% 1PA ι

U l * T F A / , n H 0 H

Solvent B:^ 25% ίΡΑ ( » 0 1 % T F A l , n H O H

0 2 0

0 1 5

010

0 05

New tools for protein separations. new HPLC column technology from Bio-Rad gives researchers a number of worthwhile alternatives for protein separations. For example, here are three separations of the same sample, using three different approaches — high performance gel filtration, our new high performance ion exchange column, and our new Hi-Pore™ reverse phase column. Looking at those separations, it's appar­ent that these HPLC columns provide fast separations with excellent resolution. Mot only that, certain compounds appear in the ion exchange and reverse phase separations that don't reveal themselves in the gel filtration separation. HPLC lets you "read between the peaks" with both speed and precision. These new columns complement rather than compete with each other. Our HPLC ion exchange column for analysis of ioniz-able biochemical compounds is an excellent complement to our Bio-Sil gel filtration column. By using all three columns, better characterization of proteins (including large ones) may be obtained. As always Bio-Rad is prepared to assist you in plotting an effective strategy for your pro­tein separations, using any or all of these techniques. Qive our applications staff a call, or contact us at

BIO-RAD Bio-Rad Laboratories — Chemical Division 2200 Wright Avenue Richmond, CA 94804 (415) 234-4130 Also in Rockville Centre, N.Υ., Australia, Austria, Canada, England, Germany, Italy, Japan, The Metherlands, and Switzerland.

CIRCLE 29 ON READER SERVICE CARD

ANALYTICAL CHEMISTRY, VOL. 55, NO. 12, OCTOBER 1983 · 1167 A