Laboratory Techniques in Biochemistry and Molecular Biology 5

599 Pages·1976·24.02 MB·english

by  T.S. Work and E. Work (Eds.)

Laboratory techniques in biochemistry
and molecular biology
5
LABORATORY TECHNIQUES
IN BIOCHEMISTRY AND
MOLECULAR BIOLOGY
Volume 5
Edited by
T.S. WORK N.I.M.R., Mill Hill, London
-
E. WORK Imperial College, London
-
Advisory board
G. POPJAK - U.C.L.A.
S. BERGSTROM - Stockholm
K. BLOCH - Haroard University
P. SIEKEVITZ - Rockejeller University
E. SMITH - U.C.L.A.
E.C. SLATER -Amsterdam
NORTH-HOLLAND PUBLISHING COMPANY - AMSTERDAM OXFORD
AMERICAN ELSEVIER PUBLISHING CO., INC. - NEW YORK
Part I Brian W. Fox
TECHNIQUES OF
SAMPLE PREPARATION OF
LIQUID SCINTILLATION COUNTING
Part I1 P.G. Righetti and J.W. Drysdale
ISOELECTRIC
FOCUSING
1976
NORTH-HOLLAND PUBLISHING COMPANY - AMSTERDAM OXFORD
AMERICAN ELSEVIER PUBLISHING CO., INC. -NEW YORK
0 1976 North-Holland Publishing Companj)
All rights reserved. No part of this publication may be reproduced, stored in a retrieval
system, or transmitted, in any form or by any means. electronic, mechanical, photocopyiny,
recording or otherwise, without the prior permission of the copyright owner.
ISBN North-Holland - series: 0 7204 4200 I
- volume 5 :0 7204 421 9 2
ISBN American Elseilier: 0 444 11216 2
Published by:
NORTH-HOLLAND PUBLISHING COMPANY - AMSTERDAM ' OXFORD
Sole distributors for the U.S.A. and Canada:
AMERICAN ELSEVIER PUBLISHING COMPANY, INC.
52 VANDERBILTA VENUE, NEW YORK, N.Y. 10017
Printed in The Netherlands
Editor’s preface
Progress in research depends upon development of technique. No
matter how important the cerebral element may be in the planning of
experiments, a tentative hypothesis cannot be converted into an
accepted fact unless there is adequate consciousness of the scope and
limitation of existing techniques; moreover, the results may be mean-
ingless or even positively misleading if the technical ‘know how’ is in-
adequate.
During the past ten or fifteen years, biochemical methods have
become specialized and sophisticated to such a degree that it is now
difficult for the beginner, whether undergraduate, graduate or special-
ist in another field, to grasp all the minor but important details which
divide the successful from the unsuccessful experiment. In order to
cope with this problem, we have initiated a new series of Laboratory
Mhnuals on technique. Each manual is written by an expert and is
designed as a laboratory handbook to be used at the bench.
It is hoped that use of these manuals will substantially reduce or
perhaps even remove that period of frustration which so often precedes
the successful transplant of a specialized technique into a new en-
vironment. In furtherance of this aim, we have asked authors to place
special emphasis on application rather than on theory; nevertheless,
each manual carries sufficient history and theory to give perspective.
The publication of library volumes followed by pocket paperbacks is an
innovation in scientific publishing which should assist in bringing
these manuals into the laboratory as well as into the library. In under-
V
VI EDITORS’ PREFACE
taking the editing of such a diverse series, we have become painfully
conscious of our own ignorance but have been encouraged by our
board of advisers to whom we owe many valuable suggestions and, of
course, by our authors who have co-operated so willingly and have so
patiently tolerated our editoral intervention.
T.S.& E. Work
Editors
Contents of parts I and I1
PART I
TECHNIQUEOSF SAMPLE PREPARATION FOR LIQUIDS CINTILLATION COUNTING.
Brian W . Fox . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
List of abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . 10
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
Chapter 1. General principles of liquid scintillation spectrometry . . . . . . . 13
Chapter 2 . Counting systems . . . . . . . . . . . . . . . . . . . . . . 41
Chapter 3. Preprocessing techniques: general aims and criteria . . . . . . . . 86
Chapter 4 . Animal tissue processing . . . . . . . . . . . . . . . . . . . 107
Chapter 5 . Botanical aspects . . . . . . . . . . . . . . . . . . . . . . . . 142
Chapter 6. Cell cultures . . . . . . . . . . . . . . . . . . . . . . . . 149
Chapter 7. Extracts and chromatographic eluates . . . . . . . . . . . . . 161
Chapter 8. Macromolecules . . . . . . . . . . . . . . . . . . . . . . . 173
Chapter 9 . Electrophoresis, centrifugation and chromatography on solid supports 184
Chapter 10. Inorganic applications . . . . . . . . . . . . . . . . . . . . 199
Chapter 11. Quench correction methods, multiple isotope counting and data
evaluation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207
Chapter 12 . Geophysics and archaeology . . . . . . . . . . . . . . . . . 244
Chapter 13 . Miscellaneous applications and future prospects . . . . . . . . 254
Appendix I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264
Appendix I1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 271
Appendix I11 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 280
Appendix IV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 284
Appendix V . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 289
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 291
Subject index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 209
VII
VIIl CONTENTS OF PARTS I AND 11
PART 11
ISOELECTRICF OCUSINPG..G . Righetti and J.W. Drysdale . . . . . . . . . . 335
Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 337
Chapter 1 . Theory and fundamental aspects of IEF . . . . . . . . . . . . 341
Chapter 2. Preparative IEF . . . . . . . . . . . . . . . . . . . . . . . 377
Chapter 3 . Analytical IEF . . . . . . . . . . . . . . . . . . . . . . . 419
Chapter 4 . General experimental aspects . . . . . . . . . . . . . . . . . 5 01
Chapter 5 . Applications of IEF . . . . . . . . . . . . . . . . . . . . . 527
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 573
Subject index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 587
TECHNIQUES OF
SAMPLE PREPARATION FOR
LIQUID SCINTILLATION
COUNTING
Brian W. Fox
Paterson Laboratories,
Christie Hospital & Holt Radium Institute,
Manehester, England