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Lab 13
Protein Identification: Western blot
20
Biology
Undergraduate 3
04/30/2012

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Term
Who started 'blotting'?
Definition
Edward Southern
Term
The scientist use what as a salt solution?
Definition
NaOH
Term
The salt solution is used to transfer _____ of _____from an _____ gel to a thin piece of _____.
Definition
Segments, DNA, agrose, nitrocellulose paper.
Term
Western blots identifies what?
Definition
Specific proteins in electrophoresis gels.
Term
Western blots use ________ methods to identify particular proteins.
Definition
Immunocytochemical
Term
Alternative name for western blotting is ...
Definition
immunoblotting.
Term
What are the antibodies produced by the B lymphocytes?
Definition
Gamma Globulin proteins, IgG
Term
What do the proteins from the B Lymphocytes do?
Definition
They bind to the antigens which are foreign chemicals introduced during an infection.
Term
Which parts do do the antibodies recognize?
Definition
epitopes, which stretch 5-10 amino acids.
Term
How do scientists begin to produce an antibody?
Definition
Injecting a known protein into the bloodstream of a mammal. Then the antibodies are produced in the rabbit.
Term
Normally researcher must do what to the rabbit to create a supply of antibiotics?
Definition
Inject it with anitgens.
Term
Since the human albumin has many possible epitopes the final collected fraction will actually contain...
Definition
a large number of different antibodies that will bind at different epitopes.
Term
The antibody is called ______, not monoclonal
Definition
polyclonal.
Term
Once you have an antibody, it will bind to a particular protein. This is a wonderful tool to...
Definition
be able to detect proteins on a western blot or to identify a particular cell location of a protein.
Term
What will be added to produce colors?
Definition
Flourecent dies like diaminobenzoic acid (DAB) or horseradish peroxidase (HRP).
Term
How are the dies coupled?
Definition
They are coupled to the primary antibody, or attached to a secondary antibody which binds to the primary antibody.
Term
Besides determining a location of a particular protein on a cell, antibodies can also be used to study...
Definition
evolutionary relationships among organisms.
Term
The aspects of immunocytochemistry relies on the antibodies to recognize...
Definition
unique amino acid sequences on proteins.
Term
In today's experiment we will compare the relatedness between
Definition
Albumin, transferrin and gamma globulins between cow, sheep, goat, horse, chicken. By comparing the electrophoresis profiles.
Term
By using an antibodies produced against cows and sheep, you will test antibodies against the different proteins on a _____ ___ identifying negligible ____ ___ differences in each proteins.
Definition
Western Blot, Amino acid
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