Antibodies Flashcards
What are antibodies?
Proteins made by immune cells that bind to foreign antigens and target them for destruction
What are the 2 regions of an antibodies?
Variable and constant
What does the variable region of an antibody do?
Is different between antibodies and binds to the antigen
What does the common region of an antibody do?
Stays the same between antibodies
Why are antibodies useful for research?
They’re extremely specific against an antigen and can be created in billions of forms. Used in numerous techniques (fluorescence microscopy, westerns, IPs)
What is a problem with using antibodies in research?
Expensive and can cross react and bind to the wrong thing
Why are antibodies useful for therapy?
For medicinal purposes, anticancer drugs. Work really well and have fewer side effects
How do you create an antibody?
Inject the protein of interest into an animal and collect the antiserum, then purify the antibody with affinity chromatography
What are polyclonal antibodies?
Different versions of an antibody that recognize the same antigen at different epitopes
What are monoclonal antibodies?
Antibodies that only recognize one epitope of the antigen
What type of antibodies do you get from injecting an animal and collecting its blood?
Polyclonal
What is an advantage of using polyclonal antibodies?
Increases the chance that at least something will bind to the protein of interest
How do you make monoclonal antibodies?
Inject a mouse with the protein of interest and use its spleen. Take those spleen cells and fuse them to cancer cells to create hybridomas. Culture those and they will keep producing antibodies forever
What is direct antibody use?
The fluorescent probe is attached directly to the primary antibody
What is indirect antibody use?
The primary antibody binds to the antigen, and the labelled secondary antibody binds to the first one
