Cells and Proteins 1a Flashcards
What is a risk
likelihood of harm arising from exposure to a hazard
Production of a standard curve can
determine the concentration of an unknown
Buffer function
allows pH to be kept constant
a centrifuge separates substances by…
density
Paper and thin layer chromatography can be used…
for separating different substances such as amino acids and sugars
what is affinity chromatography
- a solid matrix or gel column is created with specific molecules bound to the matrix or gel
- soluble targe proteins in a mixture, with a high affinity for these molecules. become attached to them as the mixture passes down the column
- Other non-target molecules with a weaker affinity are washed out
What is gel electrophoresis
- used to separate proteins and nucleic acids
- charged macromolecules move through an electric field applied to a gel matrix
How do native gels separate proteins by
by shape, size and charge
- does not denature protein
- also done by electric current
SDS-PAGE
- separates molecules by size alone
- gives all proteins an equally negative charge and denatures protein
- uses electric field
Isoelectric Point
- Molecules can be separated from a mixture using their iso electric points
- it is the pH which a soluble protein has no net charge and the protein will precipitate out the solution
- if the solution is buffered to a specific pH, only the proteins that have an IEP of that pH will precipitate
Proteins can also be separated using their IEPs in gel electrophoresis
Soluble proteins can be separated using an electric field and a pH gradient. a protein stops migrating through its gel at its IEP in the pH gradient as there is no net charge
What are immunoassay techniques used for
to detect and identify specific proteins. Uses stocks of monoclonal antibodies.
what are monoclonal antibodies
antibodies with the same specificity
An antibody specific to the protein antigen is linked to a….
chemical label
What is Western blotting technique
- done after SDS-PAGE gel electrophoresis
- The gel is run and then the separated proteins from the gel are transferred onto a solid medium
