Methods in Enzymology Volume 184 Avidin-Biotin Technology Meir Wilchek Edward A . Bayer CONTRIBUTORS TO VOLUME 184 x i PREFACE XVi i VOLUMES IN SERIES XiX Section I. Introduction 1. Reflections FREDERIC M. RICHARDS 3 2. Introduction to Avidin-Biotin Technology MEIR WILCHEK AND EDWARD A . BAYER 5 3 . Applications of Avidin-Biotin Technology : Liter- MEIR WILCHEK AND ature Survey EDWARD A . BAYER 14 Section II . Biotin-Binding Proteins 4. Biotin-Binding Proteins : Overview and Prospects EDWARD A . BAYER AN D MEIR WILCHEK 49 5. Avidin and Streptavidin N. MICHAEL GREEN 51 6 . Nonglycosylated Avidin YAFFA HILLER , EDWARD A . BAYER, AND MEIR WILCHEK 68 7 . Cloning and Expression of Avidin in Escherichia GYAN CHANDRA AND call JOHN G . GRAY 70 8 . Isolation and Properties of Streptavidin EDWARD A . BAYER, HAYA BEN-HUR, AND MEIR WILCHEK 8 0 9. Crystal Forms of Avidin ODED LIVNAH AND JOEL L . SUSSMAN 9 0 10. Nonavidin Biotin-Binding Proteins KRISHNAMURTI DAKSHINAMURT I AND JASBIR CHAUHAN 9 3 li . Biotinidase BARRY WOLF , JEANNE HYMES, AND GREGORY S . HEARD 103 12. Monoclonal Antibody to Biotin KRISHNAMURTI DAKSHINAMURTI AND EDWARD S. RECTOR 11 1 Section III. General Methodology A . Preparation of Biotin Derivatives 13. Biotin-Containing Reagents MEIR WILCHEK AN D EDWARD A . BAYER 123 14. Protein Biotinylation EDWARD A . BAYER AN D MEIR WILCHEK 13 8 15 . Enzymatic C-Terminal Biotinylation of Proteins ALEXANDER SCHWARZ , CHRISTIAN WANDREY , EDWARD A . BAYER, AN D MEIR WILCHEK 160 16. Antibodies Biotinylated via Sugar Moieties DANIEL J. O'SHANNESSY 162 17 . 2-Iminobiotin-Containing Reagent and Affinity BARBARA FUDEM-GOLDI N Columns AND GEORGE A . ORR 167 B. Preparation of Avidin and Streptavidin Derivative s 18. Avidin- and Streptavidin-Containing Probes EDWARD A . BAYER AN D MEIR WILCHEK 17 4 19. Phycobiliprotein-Avidin and Phycobiliprotein- ALEXANDER N . GLAZE R Biotin Conjugates AND LUBERT STRYER 18 8 20. Monovalent Avidin Affinity Columns RONALD A . KOHANSKI AN D M. DANIEL LANE 19 4 21. Prevention of Nonspecific Binding of Avidin RAYMOND C. DUHAMEL AN D JAMES S. WHITEHEAD 201 C . Assays for Avidin and Bioti n 22. Enzymatic and Radioactive Assays for Biotin, Av- ERNEST V. GROMAN , idin, and Streptavidin JEFFREY M . ROTHENBERG , EDWARD A . BAYER, AN D MEIR WILCHEK 208 23. Colorimetric Enzyme Assays for Avidin and EDWARD A . BAYER , Biotin HAYA BEN-HUR, AND MEIR WILCHEK 21 7 24 . Sequential Solid-Phase Assay for Biotin Based on DONALD M . MOCK 224 125 I-Labeled Avidin 25 . Fluorometric Assay for Avidin-Biotin Interaction DONALD M . MOCK AND PAUL HOROWITZ 234 Section IV. Applications A. Isolation and Purificatio n 26 . Isolation of Biologically Active Compounds : A MEIR WILCHEK AN D Universal Approach EDWARD A . BAYER 24 3 27 . Isolation and Characterization of Hormone Re- FRANCES M . FINN AND ceptors KLAUS HOFMANN 244 28 . Biotinylation of Peptide Hormones : Structural WALTER NEWMAN , Analysis and Application to Flow Cytometry L . DAWSON BEALL, AN D ZAFAR I. RANDHAWA 275 29 . Purification of Gonadotropin-Releasing Hormone ELI HAZUM 285 Receptor s 30 . Biotinylestradiol for Purification of Estrogen Re- GERARD REDEUILH , ceptor CLAUDE SECCO, AND ETIENNE-EMILE BAULIEU 292 31 . One-Step Immunoaffinity Purification of Trans- EDWARD A. BAYER AND ferrin MEIR WILCHEK 30 1 32 . Lectin-Mediated Isolation of Cell Surface Glyco- GEOFFREY M . W . COOK AN D proteins J . WILLIAM BUCKIE 304 33 . Immunoselective Cell Separation JAN WORMMEESTER , FRANK STIEKEMA, AND CORNELIS DE GROOT 31 4 34. Isolation and Analysis of Splicing Complexes PAULA J . GRABOWSKI 31 9 35 . Z-DNA Affinity Chromatography RICHARD FISHEL , PAUL ANZIANO, AN D ALEXANDER RICH 32 8 B . Localization 36. Affinity Cytochemistry MEIR WILCHEK, EDWARD A . BAYER, AN D EHUD SKUTELSKY 340 37 . Double Fluorescence Immunolabeling Studies ANNE H . DUTTON , MARGIE ADAMS, AN D S . J . SINGER 343 38 . Detection of Cell Surface Components Using Flu- ARTHUR J . SYTKOWSKI 35 3 orescent Microsphere s 39. Immunochemistry SU-MING Hsu 35 7 40 . Immunocytochemical Detection of Human T- and ROGER A . WARNKE AN D B-Cell Antigens LAWRENCE M . WEISS 363 41 . Ultrastructural Localization of Retinal Photore- DAVID S. PAPERMASTER , ceptor Proteins BARBARA G . SCHNEIDER, AND IZHAK NIR 370 42. Visualization of Intracellular Trafficking of Pro- RANDAL E . MORRIS AND teins CATHARINE B . SAELINGER 379 43 . Localization of Lectin Receptors RICHARD M . PING 388 44 . Localization of Gonadotropin-Releasing Hormone GWEN V . CHILDS 395 Receptor s 45 . Localization of Ganglioside GMI with Biotinylated HIROAKI Asou 405 Choleragen 46 . Visualization of Adenosinetriphosphatase Site of KAZUO SUTOH 409 Myosin Using a Photoreactive Adenosine 5'-Di- phosphate Analog C . Protein Blotting 47 . Analysis of Proteins and Glycoproteins on Blots EDWARD A . BAYER, HAYA BEN-HUR, AND MEIR WILCHEK 41 5 48 . Direct Labeling of Blotted Glycoconjugates EDWARD A . BAYER , HAYA BEN-HUR, AND MEIR WILCHEK 42 7 49 . Identification of Leukocyte Surface Proteins WALTER L . HURLEY AN D EVE FINKELSTEIN 42 9 50 . Staining of Proteins on Nitrocellulose Replicas WILLIAM J . LAROCHELLE AN D STANLEY C. FROEHNER 43 3 51 . Streptavidin-Enzyme Complexes in Detection of CHRISTINE L . BRAKEL , Antigens on Western Blots MARK S . BROWER, AN D KIMBERLY GARRY 43 7 52 . Luminescent Detection of Immunodot and West- MERLIN M . L . LEONG em Blots AND GLYN R . Fox 442 53 . Identification of Calmodulin-Binding Proteins MELVIN L . BILLINGSLEY , JOSEPH W . POLLI , KEITH R . PENNYPACKER, AND RANDALL L . KINCAID 45 1 D. Immunoassay s 54 . Avidin-Biotin Mediated Immunoassays : Over- MEIR WILCHEK AND view EDWARD A . BAYER 467 55 . Avidin-Biotin System in Enzyme Immunoassays THE` RESE TERNYNCK AND STRATIS AVRAMEAS 469 56. Two-Site and Competitive Chemiluminescent Im- CHRISTIAN J . STRASBURGE R munoassays AND FORTUNE KOHEN 48 1 57 . Biotinylated Protein A in Immunoassay IKUO TAKASHIMA , NOBUO HASHIMOTO, AN D HSHI-CHI CHANG 49 7 58 . Slide Immunoenzymatic Assay for Human Im- EVERLY CONWAY DE MACARIO , munoglobulin E ROBERT J . JOVELL, AND ALBERTO J . L . MACARIO 501 59 . Quantification of Carcinoembryonic Antigen in CHRISTOPH WAGENER, Serum and Analysis of Epitope Specificities of LUCIA WICKERT, AN D Monoclonal Antibodies JOHN E . SHIVELY 507 60 . Selective Precipitation of Biotin-Labeled Antigens CHRISTOPH WAGENER, or Monoclonal Antibodies by Avidin for Deter- ULRICH KRÜGER, AN D mining Epitope Specificities and Affinities in JOHN E . SHIVELY 518 Solution-Phase Assay s 61 . Immunoassays for Diagnosis of Infectious Dis- ROBERT H . YOLKEN 529 ease s 62 . Enzyme-Linked Immunosorbent Assay WILLIE F. VANN , ANN SUTTON, AND RACHEL SCHNEERSON 537 63. Rapid Detection of Herpes Simplex Virus LATA S . NERURKAR 54 1 64 . Sensitive Immunoassay for Human Immunodefi- KENNETH R . HUSKINS , ciency Viral Core Proteins KEVIN J . REAGAN , JOHN A . WEHRLY, AND JEANNE E . NEUMANN 556 E. Gene Probe s 65. Gene Probes MEIR WILCHEK AN D EDWARD A. BAYER 560 66 . Biotinylated Nucleotides for Labeling and Detect- LEONARD KLEVAN AN D ing DNA GULILAT GEBEYEHU 561 67 . Biotinylated Psoralen Derivative for Labeling Nu- COREY LEVENSON , cleic Acid Hybridization Probes ROBERT WATSON, AN D EDWARD L. SHELDON 57 7 68 . Chemically Cleavable Biotin-Labeled Nucleotide TIMOTHY M . HERMAN AND Analogs BARBARA J . FENN 58 4 69 . Preparation and Uses of Photobiotin JAMES L . MCINNES , ANTHONY C . FORSTER , DEREK C . SKINGLE, AN D ROBERT H . SYMONS 588 70. Modification of Cytosine Residues on DNA RAPHAEL P. VISCIDI 600 71. Direct Incorporation of Biotin into DNA MEIR WILCHEK , JEFFREY M . ROTHENBERG , AVI REISFELD, AN D EDWARD A . BAYER 608 72. Colorimetric-Detected DNA Sequencing STEPHAN BECK 61 2 F. Composite and Special Applications 73. Composite Applications MEIR WILCHEK AND EDWARD A . BAYER 61 8 74. Third Component of Human Complement: C3 MELVIN BERGER 61 9 75 . Purification and Characterization of Membrane JEAN-PIERRE KRAEHENBUH L Proteins AND CLAUDE BONNARD 62 9 76 . Facilitated Cell Fusion for Hybridoma Production MARY K . CONRAD AND MATHEW M . S . Lo 64 1 77 . Complex Neoglycoproteins MING-CHUAN SHAG , LING-MEI CHEN, AND FINN WOLD 65 3 78. Probing ß-Adrenergic Receptors KATHRYN E . MEIER AN D ARNOLD E . RuoHo 66 0 79 . Probing Adenosine Receptors Using Biotinylated KENNETH A . JACOBSON 66 8 Purine Derivative s AUTHOR INDEX 67 3 SUBJECT INDEX 705.