elisa Flashcards
Your body has three layers of defense against pathogens (disease-causing microorganisms). What are they?
*Natural Barriers
*Non-specific immune response
*Specific immune response
Is natural barriers first, second, or third line of defense?
What is it made up of?
What happens if there is a tear or break in the barriers?
*Body’s first line of defense
*Made up of the skin and mucous membranes, as well as bodily secretions such as saliva, sweat and tears
*A tear or break in these barriers provides an entry point for pathogens
What is another name for non-specific immune response?
What does it generally respond to? What does it involve?
*AKA innate immunity
*Involves general responses to infection, such as fever and inflammation
*Involves white blood cells that do not target specific pathogens, such as NK cells and macrophages.
What are specific blood cells called and what is their function? What do B cells make? Antibody recognize and bind to a specific antigen on a pathogen why?
*Here, specific blood cells called Lymphocytes (B cells and T cells) pick up a piece of a pathogen that it can learn to recognize, called an antigen
*Antigens are typically proteins or parts of proteins found on the surface of a pathogen or foreign substance
*B cells make antibodies (aka immunoglobulins, or Igs) designed to target this antigen
*Antibodies recognize and bind to a specific antigen on a pathogen, tagging it for destruction by the immune system
What are antigens?
*Antigens are typically proteins or parts of proteins found on the surface of a pathogen or foreign substance
What are antibodies and their function? Which cells make antibodies?
(aka immunoglobulins, or Igs) designed to target this antigen
*Antibodies recognize and bind to a specific antigen on a pathogen, tagging it for destruction by the immune system
B cells
Why are antibodies important?
Needed for immune memory
-Can be used for treatment for illnesses
-Used as a tool for to recognize a protein of interest
*Western Blot
*Flow-cytometry
*Fluorescent microscopy
*ELISA
*Immunoprecipitation
Which tools identify antibodies
Western Blot
*Flow-cytometry
*Fluorescent microscopy
*ELISA
*Immunoprecipitation
What does ELISA stand for?
*Enzyme-linked Immunosorbent Assay
What is ELISA
a plate-based assay technique designed for detecting and quantifying soluble substances such as peptides, proteins, antibodies, and hormones.
*An antigen and antibody are mixed together, and the interaction is detected via a reporter attached to the antibody
The most crucial element of an ELISA is a highly specific antibody-antigen interaction
What are the 4 types of ELISA
*Direct
*Indirect
*Sandwich
*Competitive
Direct ELISA
*A plate is coated with an antigen.
*A conjugate antibody (an antibody with a reporter attached) specific to the antigen is then added
*Pros: Easy and straight forward
*Cons: Can get expensive fast
Indirect ELISA
A plate is coated with an antigen.
*An antibody specific to the antigen is applied (this is the primary antibody)
*A secondary conjugate antibody that recognizes the Fc region of the primary antibody is applied
Sandwhich ELISA
*A plate is coated with an antibody (this is the capture antibody)
*A sample containing your antigen is applied and binds to the capture antibody
*An antibody specific to the antigen is applied (this is the primary antibody)
*A secondary conjugate antibody that recognizes the Fc region of the primary antibody is applied
competitive ELISA
*measures the concentration of an antigen by detection of signal interference.
*The sample antigen competes with a reference antigen for binding to a specific amount of labeled antibody
The more sample antigen there is, the less binding of the inhibitor to the labelled antibody, and therefore the lower the signal
