biochem lab part 5w

what does in vivo mean

research done on a living organism

what does in vitro mean

research done in a laboratory dish or test tube

what are the two types of traditionally produced antibodies

polyclonal antibodies and monoclonal antibodies

what are polyclonal antibodies

generated by immunizing an animal (usually a rabbit, goat, or sheep) and obtaining serum. the product is antiserum that can be used directly or the antibodies can be purified from it.

using polyclonal bodies in diagnostic tests are too _____

variable

what is a monoclonal antibody

B cells are infused with tumor-like immortal cells that generate hybrid cells which can be cultured indefinitely and purified from a growth medium with almost no batch to batch variability

what is the basis of genetically engineering antibodies

Recombinant DNA technology can be used to produce antibodies that look human to the human immune system and so can be used as therapeutic agents in people.

what is hybridoma immortalization

Recombinant DNA technology allows the antigen recognition site from a known mouse monoclonal antibody to be camouflaged within a human antibody by combining part of the mouse gene with the human antibody gene. Bacteria transformed with this DNA are capable of producing humanized monoclonal antibodies indefinitely

what is the basis of phage display

inserting shuffled antibody genes from billions of human B cells into the genomes of bacteriophage lambda (bacteriophages, or phages, are viruses that infect bacteria; lambda phage is a specific species of phage), so that the lambda phages display the binding sites from human antibodies on their surfaces. can be cloned into a human antibody gene and transformed into bacteria, then large amounts of antibody can be produced for therapeutic use

what are used in diagnosis and research as labeling tools

antibodies. as labels they have to be made visible so antibodies are covalently linked

how can antibody targets or antigens be detected directly

by labeling the antibody specific for the antigen and looking for signal

what is indirect detection

uses polyclonal secondary antibodies to recognize primary antibodies, then primary antibodies bind specifically to the antigen and the secondary antibody binds specifically to the primary antibody. means that only one type of enzyme linked secondary antibody is needed to visualize all antibodies produced in one type of animal

how are secondary antibodies produced

by injecting the antibodies of one animal into a different species of animal

what is immunostaining

localizes antigens in organelles, cells, tissues, or whole organisms, and can also be used to distinguish one cell type from another. can be used to distinguish cancer cells from normal cells

what is fluorescence activated cell sorting (FACS)

special application of immunostaining in which a population of cells is stained with a fluorescently labeled antibody and then physically separated into labeled and unlabeled cells, then fluorescent labels are detected

what is immunoblotting

tells us about a protein’s size and relative abundance in a given sample. probed with primary antibody that is specific for the protein of interest, then an enzyme linked substrate

what is dot blotting

modification of immunoblotting in which a sample is spotted onto a membrane directly, used for the rapid screening of a large number of samples. provides no information abt size of the protein

what do dipstick tests for illegal drug test include

marijuana, cocaine, and methamphetamines

what do dipstick tests for infectious agents test include

HIV, plague, E. coli O157, and Legionella

what type of test is a dipstick test

immunochromatography assays

what is a dipstick test you can get in a pharmacy

pregnancy test