Lab Techniques! Flashcards
Hazards in the lab
Toxic or corrosive chemicals
Flammable substances
Pathenogenic organisms
Mechanical Equipment
Risk definition
Liklehood of harm arising from exposure to hazard
Risk assessment=Identifying control measures to minimise risk
Control Measures
Using appropriate handling techniques protective clothing and equipment and aseptic technique
Centrifuge
To separate substances of differing density. More dense components settle in pellet less dense remain in the supernatant.
Keep cold to keep proteins from denaturing
Paper and thin layer chromatography
Can be used for separating diff substances such as amino acids and sugar
Speed each solute travels along chromatogram depends on differing
solubility
How affinity chromatography seperates proteins
-Solid matrix has specific molecules bound
- Soluble target proteins have high affiinity and attatch to them as mizture passes down column
- Other non target molecules are washed out
Gel Electrophoresis
-Charged macromolecules move through electric field applied to gel matrix
-Native gels separate proteins
- Do not denature molecule
How are proteins separated in
Gel Electrophoresis
Shape, Size, Charge.
Proteins seperated by … in
SDS Page
Size Alone
Gives all molecules equally neg charge and denatures them
How can
Proteins be separated by a mixture
Using their isoelectric points
IEP’s
What is the
Isoelectric point
The pH at which a soluble protein precipitates out of solution.
Monocolonal antibodies
Same specificity
Detecting proteins using antibodies
- ## An antibody specific to antigen linked with chemical label
Labels often are
Reporter enzyme producing colour change
- chemiluminescence
- flourescence
Western Blot
-Used after SDS Page electrophoresis
-Separated proteins from gel are transferred to a solid medium
-Proteins can be identified with specific antibodies with reporter enzymes attatched
