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10 Cards in this Set
- Front
- Back
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Why are analytical protein techniques used?
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Measuring purity of proteins, measuring the concentration/amount of proteins, diagnosis, mechanistic studies, determining molecular weight and structural studies (which also use X-Ray diffraction techniques)
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PAGE stands for......
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Polyacrylamide Gel Electrophoresis
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The sheets of 'gel' employed in PAGE are composed of.....
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Acrylamide (0.8%)
Bis-acrylamide (0.2%) |
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What factors affect the degree of separation in PAGE?
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- Amount of Cross-linking (which relates to Bis-Acrylamide concentration)
- Charge Density -(Charge/Volume) Smaller molecules travel faster |
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What does SDS stand for?
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Sodium-Dodecyl-Sulphate
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What is the main chemical property of SDS and why is it useful?
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SDS is an ampipathic molecule. Causes protein structures to be denatured (reducing) and coated in a uniform, net charge. Note that quaternary structure proteins are separated into their separate sub-units.
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A patient has a suspected Myocardial Infarction. How might you confirm this?
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Perform SDS-Page.
Different isoforms of Creatine Phosphokinase exist, one is found only in cardiac muscle. Leaks into blood serum when MI occurs. Different molecular weight. Calibrate using markers. Take sample of serum and perform SDS Page. |
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A post-transplant patient's blood serum is tested using PAGE. It is scanned to quantify the levels of gammaglobulins present. What results do you expect to see?
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Very low levels of gammagobulins. They will be immunosupressed from immunosupressant medication.
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Why is it possible to use PAGE to diagnose lipidaemias?
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LDLs, VLDLs and HDLs have different charges.
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Why would PAGE not be useful to compare chylomicron levels?
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Chylomicrons do not migrate from the origin when a voltage is applied accross the gel.
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