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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Positive sample
negative sample
5. Unbound labeled antibody is washed away and a colorimetric substrate is added. Draw this below
Positive sample
negative sample
6. The enzyme cleaves the substrate causing a color change of the substrate solution. The intensity of
the color is quantified using a spectrophotometer and is proportional to the amount of antibody
how to do that?).
present in the sample. (Obviously, a standard curve would have to be generated. Could you think of
Positive sample
negative sample
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Transcribed Image Text:Positive sample negative sample 5. Unbound labeled antibody is washed away and a colorimetric substrate is added. Draw this below Positive sample negative sample 6. The enzyme cleaves the substrate causing a color change of the substrate solution. The intensity of the color is quantified using a spectrophotometer and is proportional to the amount of antibody how to do that?). present in the sample. (Obviously, a standard curve would have to be generated. Could you think of Positive sample negative sample
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.
expand button
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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Can you do the first part?

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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.
expand button
Transcribed Image Text: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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Follow-up Questions
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Follow-up Question

Can you do the first part?

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Follow-up Question
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.
expand button
Transcribed Image Text: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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