Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN: 9780134580999
Author: Elaine N. Marieb, Katja N. Hoehn
Publisher: PEARSON
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The purpose of ELISAS is to detect specific protein antigens using antibodies. In laboratory medicine,
ELISAS are commonly used to screen serum samples from animals to detect antibodies to a specific
pathogen. In the research environment, ELISAS can be modified to detect a protein of interest.
Method:
1. 96 well plates are coated with an antigen. Sites unoccupied by the antigen are blocked with a
blocking buffer to prevent non-specific binding of sample. Draw this below:
2. Serum samples are added and incubated to allow specific antibodies to bind to specific antigen.
Antibodies that do not recognize and bind specifically to the antigen remain unbound. Draw this
below:
Positive sample
Negative sample
3. Non-specific antibodies that do not bind to the antigen are washed away. Draw this below.
Positive sample
Negative sample
4. An enzyme labeled (e.g. alkaline phosphatase or horseradish peroxidase) secondary antibody is
incubated with the samples and binds tightly to sample antibody. Draw this below.
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Transcribed Image Text:The purpose of ELISAS is to detect specific protein antigens using antibodies. In laboratory medicine, ELISAS are commonly used to screen serum samples from animals to detect antibodies to a specific pathogen. In the research environment, ELISAS can be modified to detect a protein of interest. Method: 1. 96 well plates are coated with an antigen. Sites unoccupied by the antigen are blocked with a blocking buffer to prevent non-specific binding of sample. Draw this below: 2. Serum samples are added and incubated to allow specific antibodies to bind to specific antigen. Antibodies that do not recognize and bind specifically to the antigen remain unbound. Draw this below: Positive sample Negative sample 3. Non-specific antibodies that do not bind to the antigen are washed away. Draw this below. Positive sample Negative sample 4. An enzyme labeled (e.g. alkaline phosphatase or horseradish peroxidase) secondary antibody is incubated with the samples and binds tightly to sample antibody. Draw this below.
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