Antibody Techniques

Author： Malik Vedpal S.;Lillehoj Erik P.

Publisher： Elsevier Science‎

Publication year： 2013

E-ISBN: 9781483285139

P-ISBN(Paperback): 9780124664609

P-ISBN(Hardback): 9780124664609

Subject： R1 Preventive Medicine , Health;R392 medical immunology

Language： ENG

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Description

The applicability of immunotechniques to a wide variety of research problems in many areas of biology and chemistry has expanded dramatically over the last two decades ever since the introduction of monoclonal antibodies and sophisticated immunosorbent techniques. Exquisitely specific antibody molecules provide means of separation, quantitative and qualitative analysis, and localization useful to anyone doing biological or biochemical research.

This practical guide to immunotechniques is especially designed to be easily understood by people with little practical experience using antibodies. It clearly presents detailed, easy-to-follow, step-by-step methods for the widely used techniques that exploit the unique properties of antibodies and will help researchers use antibodies to their maximum advantage.

Detailed, easy-to-follow, step-by-step protocols
Convenient, easy-to-use format
Extensive practical information
Essential background information
Helpful hints

Chapter

CHAPTER 1. ANTIBODY MOLECULES AND THE IMMUNE RESPONSE: THEORETICAL BACKGROUND

I. THE ORGANIZATION OF IMMUNE PROTECTION

II. INNATE AND ACQUIRED IMMUNITY

III. ACQUIRED IMMUNITY HAS TWO EFFERENT LIMBS

IV. PRIMARY AND SECONDARY LYMPHOID ORGANS

V. THE TEMPO AND MAGNITUDE OF IMMUNE RESPONSES

VI. CELLULAR EVENTS AND CROSS-TALK

VII. AFFINITY MATURATION

VIII. PREVENTION AND TERMINATION OF HUMORAL RESPONSES

IX. CONCLUSION

Acknowledgment

References

CHAPTER 2. ADJUVANT-FREE POLYCLONAL ANTIBODY RESPONSE MANIPULATED BY ANTIBODY-MEDIATED ANTIGEN TARGETING

I. INTRODUCTION

II. ANTIBODY-MEDIATED ANTIGEN TARGETING: AN OVERVIEW

III. METHODS

Acknowledgment

References

CHAPTER 3. HYBRIDOMAS AND MONOCLONAL ANTIBODIES

I. INTRODUCTION

II. MATERIALS

III. PROCEDURES

References

CHAPTER 4. PCR AMPLIFICATION OF ANTIBODY GENES

I. INTRODUCTION

II. METHODS

III. RESULT AND DISCUSSION

Acknowledgment

References

CHAPTER 5. LABORATORY ANIMAL CARE AND USE

I. INTRODUCTION

II. LAWS AND REGULATIONS

III. ANIMAL CARE

IV. ANIMAL USE

V. OCCUPATIONAL SAFETY AND HEALTH

References

CHAPTER 6. ADJUVANTS

I. INTRODUCTION

II. PREPARATION OF ADJUVANTS AND VACCINES

References

CHAPTER 7. LARGE-SCALE PRODUCTION OF MONOCLONAL ANTIBODIES

I. INTRODUCTION

II. FACTORS TO CONSIDER

III. LARGE-SCALE PRODUCTION

IV. CONCLUDING REMARKS

References

CHAPTER 8. ANTIBODY-BINDING BACTERIAL PROTEINS AS IMMUNOREAGENTS

I. BACTERIAL IMMUNOGLOBULIN-BINDING PROTEINS

II. IgG-BINDING PROTEINS AS IMMUNOCHEMICAL REAGENTS

III. OVERVIEW AND FUTURE DIRECTION

Acknowledgment

References

CHAPTER 9. CONJUGATING ANTIBODIES TO LIGANDS, ENZYMES, AND METALLIC PARTICLES

I. FLUOROCHROME-CONJUGATED ANTIBODIES

II. BIOTINYLATION OF ANTIBODIES AND OTHER MACROMOLECULES

III. ENZYME-LABELED ANTIBODIES

IV. ANTIBODIES CONJUGATED TO COLLOIDAL GOLD

References

CHAPTER 10. DOT IMMUNOBINDING (DIB), ENZYME-LINKED IMMUNOSORBENT ASSAY(ELISA), AND RADIOIMMUNOASSAY (RIA) FOR DETECTING PEPTIDE ANTIGENS AND SPECIFIC ANTIBODIES

I. INTRODUCTION TO SOLID-PHASE IMMUNOASSAYS AND RADIOIMMUNOASSAYS

II. DOT IMMUNOBINDING (DIB) OF AN OCTAPEPTIDE, ANGIOTENSIN II

III. ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA)

IV. RADIOIMMUNOASSAY FOR THE DECAPEPTIDE ANGIOTENSIN I

References

CHAPTER 11. IMMUNOSTAINING CELLS AND TISSUES

I. INTRODUCTION

II. CELL AND TISSUE PREPARATION

III. CONTROLS

IV. PROCEDURES

V. DISCUSSION

References

CHAPTER 12. PROTEIN IMMUNOBLOTTING

I. INTRODUCTION

II. SEPARATION OF PROTEINS BY SDS–PAGE

III. ELECTROTRANSFER OF PROTEINS

IV. IMMUNODETECTION OF IMMOBILIZED PROTEINS

V. PROCEDURES

VI. TROUBLESHOOTING

References

CHAPTER 13. FLOW CYTOMETRY AND FLUORESCENCE-ACTIVATED CELL SORTING

I. INTRODUCTION

II. INSTRUMENTATION

III. APPLICATIONS

IV. PROCEDURES

V. PITFALLS AND CONCERNS

References

CHAPTER 14. SCREENING EXPRESSION LIBRARIES WITH ANTIBODIES

I. INTRODUCTION

II. MATERIALS

III. PROCEDURES

IV. POTENTIAL PITFALLS

References

CHAPTER 15. IMMUNOAFFINITY PURIFICATION

I. INTRODUCTION

II. MATERIALS

III. PROCEDURES

References

INDEX

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