Crowther ELISA

Theory and Practice
1995
ISBN: 978-0-89603-279-8
Verlag: Springer, Berlin

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Beschreibung

7he a1m 0f EL15A: 7he0ry and Pract1ce t0 1ntr0duce the expe- menter t0 the techn0109y kn0wn the En2yme L1nked 1mmun050r6ent A55ay (EL15A). 7he 600k d1rected at w0rker5 w1th vary1n9 de9ree5 0f exper1ence, 6ut f0r th05e w1th0ut any prev10u5 c0ntact w1th the a55ay. 1mmun0a55ay5 f0rm the 6ack60ne 0f u5ed te5t5 the 5tudy 0f 1nf- d15ea5e5 t10u5 and end0cr1n0109y, and hence can 6e u5ed t01d- a9ent5 cau5at1ve t1fy 0f the extent and nature (ep1dem10109y) 0f d15ea5e, and mea5ure h0rm0ne5 at EL15A5 pr0v1de h19h1y and prec15e meth0d5 f0r the mat10n 0f parameter5, w1th the added advanta9e that they can hand1e 1ar9e num6er5 0f 5amp1e5 that may then 6e ana1y2ed rap1d1y. 7he5e a55ay5 are ver5at11e, hav1n9 6een app11ed t0 a w1de ran9e 0f e.9., v1ru5e5, 6acter1a, fun91, and pr0t020an and meta- 20an 51nce para51te5. 1971, when the effect1ve en2yme-1a6e1ed a55ay wa5 de5cr16ed (1), th0u5and5 0f have 6een pu6115hed de- w1th the 4uant1f1cat10n 0f ant19en5 and ant160d1e5 f0r re5earch and app11ed purp05e5. EL15A and re1ated a55ay5 1nv01v1n9 u5e 0f en2yme5 t0 06ta1n c010r1metr1c re5u1t5 have n0w rep1aced rad101mmun0a55ay (R1A) f0r m05t d1a9n05t1c purp05e5, 51nce the f0rmer 0ffer5 a p0tent1a1 w1th 1ncrea5ed ver5at111ty t0 a w1der 9r0up 0f Examp1e5 0f 5pec1f1c area5 t0 wh1ch EL15A5 have 6een app11ed re5earch and are: Detect10n and 0f wh01e 0r part5 0f a9ent5, e.9., typ1n9; 2. 0f a9ent5, e.9., 5u6typ1n9; d15ea5e D15cr1m1nat10n d15ea5e 1dent1f1cat10n 1. d1a9n0515 1n 5c1ent15t5.

Autoren

Autoren

Crowther, John R.

Inhaltsverzeichnis

Basic Immunology:
1. The Immune Response
2. Antigens
3. Adaptive Immunity and Clonal Selection
4. Antibodies
5. Antigenic Commonness
Basic Principles of ELISA:
1. Reaction Schemes
2. Direct ELISA
3. Indirect ELISA
4. Sandwich ELISA 5. Competition ELISA
6. Choice of Assays
Stages in ELISA:
1. Solid-Phase
2. Washing
3. Addition of Reagents
4. Incubation
5. Blocking Conditions and Nonspecific Reactions
6. Enzyme Conjugates
7. Availability of Conjugates
8. Conjugation with Enzymes
9. Development of Label
10. Stopping Reactions
11. Reading
12. Practical Problems
Theoretical Considerations:
1. Setting Up and Use of ELISA
2. What Is Already Known
3. Complexity of Problems
4. Antigenic Considerations
5. Other Techniques
6. Units
7. Making Dilutions
8. Pipets
9. Molarities
Direct ELISA:
Titration of Antigen and Antibody:
1. Learning Principles
2. Reaction Scheme
3. Basis of Assay
4. Materials and Equipment
5. Practical Details
6. Data Explained
7. Aspects of Assay Described
8. Conclusions
Indirect ELISA:
1. Learning Principles
2. Basis of Assay
3. Materials and Reagents
4. Practical
5. Use of Indirect ELISA to Titrate Antibodies
6. Use of Indirect ELISA to Determine the Positivity of Sera at Single Dilution
Use of Antibodies on Solid-Phase in Capture ELISA:
1. Use of Capture ELISA to Detect and Titrate Antigen
2. Reaction Scheme
3. Basis of Assay
4. Materials
5. Methods
6. Titration of Capture Antibody Using IgG
7. Titration of Capture Antibody when Used as Whole Serum
8. Use of Capture ELISA to Detect Antigens
9. Use of Capture ELISA to Detect and Titrate Antibodies
Competitive ELISA:
1. General Information
2. Direct Competitive ELISA for Antigen Detection and Quantification
3. Indirect Assay-Antigen Competition
4. Indirect Competition Assay for Antibody Detection
5. Indirect Assay Competition for Antibody-Detection Using a Single Dilution of Test Serum
6. Overall Conclusions on Competition Assays
Immunochemical Techniques:
1. Introduction
2. Labeling Antibodies with Enzymes
3. Coupling Antibodies to Alkaline Phosphatase
4. Preparation of Immunoglobulins
5. Immunosorbents
6. Production of Antisera

Zielgruppe

Research

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