NBSIR 86-3345 NAT'L INST. OF STAND & TECH in II III II ll ill II ii mu I III III I llll AlllOti M30b51 - - ^ X 1986 MEETING OF THE / AMERICAS BRANCH -.3 OF THE ELECTROPHORESIS SOCIETY \ % *v '•V-. TH, MARCH 26-28, 1986 PROCEEDINGS Editor: Dennis J. Reeder SPONSORED DY: U.S. DEPARTMENT OF COMMERCE National Bureau of Standards National Measurement Laboratory NATIONAL BUREAU OF STANDARDS Center for Analytical Chemistry _ Gaithersburg, MD 20899 AND THE ELECTROPHORESIS SOCIETY March 1986 QC 100 .U56 NO. 86-3345 U.S. DEPARTMENT OF COMMERCE, Malcolm Baldrige, Secretary National Bureau of Standards, Ernest Ambler, Director 1986 a. 3- N NBSIR 86-3345 i#-: A 986 MEETING OF THE / AMERICAS BRANCH . THE OF ' r.~ *~c- ELECTROPHORESIS"SOCIETY ?-£ -i 'A i : . ? V:’*- T» jr ‘ TS,„ MARCH 26-28, 1986 PROCEEDINGS Editor: Dennis J. Reeder SPONSORED DY: U.8. DEPARTMENT OP COMMERCE National Bureau of Standards National Measurement Laboratory NATIONAL BUREAU OF STANDARDS Center for Analytical Chemistry _ Galthersbu rg. M D 20899 A U THE ELECTROPHORESIS SOCIETY March 1986 U.S. DEPARTMENT OF COMMERCE, Malcolm Baldrige, Secretary National Bureau of Standards, Ernest Ambler, Director INTRODUCTION The Electrophoresis Society (America's Branch) in 1985 had the task of deciding whether to hold a national meeting in 1986, the same year as the planned international meeting in London. As the council met, the consensus was that there would be sufficient interest in a meeting that plans should be made. The authors of the plenary sessions were contacted and graciously committed to the task of writing papers for publication in the proceedings. The meeting format was agreed upon to allow maximum interactions among the participants, to allow sufficient time for plenary lecturers to give a comprehensive lecture, and to enable the attendees to have a published document for later reference. The afternoon meetings were structured to permit the greatest information exchange by arranging sessions on selected topics, moderated by session chairpersons who would facilitate the direction of the presentations. The cooperation and willingness of individuals to participate would be the key to a successful information exchange. The informal nature of the sessions was devised to allow the expert and the novice to exchange findings and details about their research. The impact of electrophoretic separations on biochemical analyses is reflected in the topics covered in this publication. Two-dimensional electrophoresis (2- DE) has matured to become an accepted tool for clinical studies. Immobilized pH gradients are becoming more reproducible and have found use as the first dimension high resolution separation step in 2-DE. The chemistry of protein detection in gels has progressed significantly to enable quantification. Although not fully understood, studies of staining mechanisms are proceeding. Nucleic acid separations in gels is a common technique, but the theoretical aspects of the separation process are just now being explored. Standardization in electrophoretic separations is addressed in relation to new materials for molecular weight, size and charge standards. The beginnings of interlaboratory standardization by round-robin studies also is reported. New matrix materials for separations are introduced . Finally, the practical applications of many techniques are reported. II ' . ACKNOWLEDGEMENTS This publication was compiled from papers and abstracts submitted for the Electrophoresis Society Meeting. I am particularly grateful to Jesse J. Edwards and Margaret C. Kline for their work in the final days of preparation. For helpful discussions, I am also indebted to Diane K. Hancock, Kathy C. Stang and Kathleen D. Kilmer DISCLAIMER The use of trade names and manufacturer's names in this publication in no way represents an endorsement of a particular product or manufacturer by the National Bureau of Standards or the Electrophoresis Society. iii . CONTENTS PLENARY PAPERS: Bruce Budowle Making Ultrathin- layer Polyacrylamide Gel Isoelectric Focusing a Reproducible Methodology John Fawcett 13 Polyacrylamide Gel Electrofocusing in pH gradients formed by Carrier Ampholytes and by Immobiline: Pro's and Con's Carl R. Merril 36 Stains for the Detection of Proteins Separated by Electrophoretic Methods Susan C. Olson and Carl R. Merril 66 Peptide Separations: Techniques Reviewed Philip Serwer 86 Viruses and Viral Components in Extracts of Infected Cells: Analysis by Agarose Gel Electrophoresis Nancy Stellwagen 10* Gel Electrophoresis of DNA * N. L. Anderson's paper on Macromolecular Identification on Two-Dimensional Gels was not received in time for publ icat ion ABSTRACTS: Stephen H. Blose 1J7 The Functions of a Protein Database Developed from Comput e r - Ana 1 v/.ed Quantitative Two-Dimensional Gels Yvon C . Chagnon Genetic Study of Marine Vertebrate and Invebrate Populations Using Ultra-Thin Isoelectric Focusing Techniques Gail Chuck and George Hug 1 J9 Detection of Hexosaminidase Isozymes by Slab Gel Isoelectric Focusing (PAG-IEF) Richard B. Cook NuFix Glyoxyl Agarose: A Versatile New Medium for Electrophoresis, Blotting and Chromatography Dale Dykes The Use of Biotinylated Human DNA Probes for Detect inr. P. FhP'.s Anne M. Gambel, Bruce Budowle, and Randall S. Murch Alpha 1-Antitrypsin (PI) Phenotyping by Ultrathin haver I socle, trio Focusing James I. Garrels and Robert Franza, Jr 143 New Development in Hardware and Software for 2D Gel Analysis . Douglas M. Gersten, E.J. Zapolski, T. J. Golab, and R.S. Ledley . 144 Automated Nucleic Acid Electrophoresis and Hybridization Analysis Robert L. Gilman • • 1^5 Electrophoretic Separation of High Density Lipoprotein Fractions E. Gombocz D. Tietz, S. Hurtt, and A. Chrambach 146 , Physical Characterization of Turnip Crinkle and Pellargonium Flower Break Viruses by Agarose Gel Electrophoresis: Applicability of Polystyrene Size Standards and of Moving Boundary Electrophoresis Buffer Systems Angelika Gorg, W. Postel, S. Gunther, and J. Weser . 147 Immobilized pH Gradients for One- and Two-Dimensional Separations. Applications Samir M. Hanash, L.J. Baier, and Rork Kuick 148 A New Prognostic Indicator in Acute Lymphoblastic Leukemia Detected by Two-Dimensional Polyacrylamide Gel Electrophoresis (2-D PAGE) Diane K. Hancock 149 Mechanistic Studies of Silver Staining William E. Heydorn, G. J. Creed, C. R. Creveling and D.M. Jacobowitz 150 Studies on Catechol-O-Methyltransferase in Rat Brain using Two- Dimensional Gel Electrophoresis Ann E. Kaplan and G.N. Gray 151 Immobiline Electrophoresis: pi Values for Proteins of LDH-4 and -5 from Control and Nitrosomethylurea-Transformed Neoplastic Cell Lines Reinhold C. Mann, B.K. Mansfield, and J. K. Selkirk 152 Parametric Versus Non-Parametric Gel Image Analysis Betty K. Mansfield, R. C. Mann, and J. K. Selkirk ...... 153 5 2 , -Diphenyloxazole (PPO) in Dimethyl Sulfoxide (DMSO) Regeneration for Re-Use in Fluorography Mark Miller and S.S. Thorgeirsson 154 Analysis of Micro -Variations in the Rate of Synthesis of Individual Polypeptides Isolated from Cultures of the Rat Hepatoma Cell Line H4-II-E as Measured by Quantitative Two-Dimensional Gel Electrophoresis V Rodger Morrison and M.G. Brown 155 Beta-Hemoglobin Polymorphism in White-Tailed Deer: Suitability and Application to Wildlife Law Enforcement James E. Myrick, M.K. Robinson, I.L. Hubert, C.J. Bell, and S. P. Caudill Preparation of Urine and Serum Samples with Centrifugal Microconcentrators and Total Protein Analysis by Ultraviolet-Difference Spectroscopy for Two-Dimensional Electrophoretic Analysis Stephen Peats, S. Nochumson, F.H. Kirkpatrick Effects of Borate on Agarose Gel Structure Kristy L. Richie and D.J. Reeder Results of an Interlaboratory Round Robin Using Isoelectric Focusing to Assess Reproducibility of Rehydratable Gels Atul Sahai and S. M. Hanash Elevated Levels of p21 ras Detected in Blast Cells from Leukemic Patients David Sammons and R. Humphreys Intermediate Range Molecular Weight Markers for SDS Gels Burton E. Sarnoff, D. Berretta and P. Todd The Effect of Calcium Ion Concentration on the Electrophoretic Mobility of Two Cultured Human Embryonic Kidney Cell Populations Cheri Seitz Advantages of Focusing Carbamylation Trains in a Flat -Bed System John R. Strahler, S. Hanash, L. Somerlott and A. Gorg Two-Dimensional Polyacrylamide Gel Electrophoresis of Lymphocyte Proteins with Immobilized pH Gradients in the First Dimension Dietmar Tietz, E. Gombocz and A. Chrambach Physical Characterization of Particles and Gel Fibers on the Basis of Non-Linear Ferguson Plots in Agarose Gel Electrophoresis Kaija H. Valkonen and D. Goldman 1 Preparative and Analytical Immobiline Electrofocus i ng of lass 111 Hepatic Alcohol Dehydrogenase Peter J. Wirth and S. S. Thorgeirsson 166 Sequential Analysis of Chemically Induced Hepatoma Development Using Two-Dimensional Electrophoresis (2-D PAGE) Virginia P. Wray 167 Foundation Experiments for Building a Protein Database Joseph Yudelson 168 Mechanism of Nickel and Silver Visualization of Protein Electrophoregrams Rashid A. Zeineh, J. Zeineh and M. Zeineh 169 Comparison of 2 -Dimensional Gel Electrophoretic Separations Using Soft Laser Scanning Densitometry with Computer Color Graphics Rashid A. Zeineh, J. Zeineh and M. Zeineh 170 Multilane Strip -Chart Printing Program for DNA Sequence Reading vii PLENARY PAPERS MAKING ULTRATHIN-LAYER POLYACRYLAMIDE GEL ISOELECTRIC FOCUSING A REPRODUCIBLE METHODOLOGY Bruce Budowle Forensic Science Research and Training Center, Laboratory Division, FBI Academy, Quantico, Virginia