Flashcards in Practical 1 - Spectrophotometry and Electrophoresis Deck (10):
Method of analysing molecules based on charge, by measuring their migration in an electric field.
Method of analysing molecules based on their spectral properties
What is absorbance?
Fraction of incident light absorbed by a solution which is measured.
Absorbance = log10 (light transmitted through blank solution/light transmitted through test solution)
At which wavelength is there maximum haemoglobin absorption?
540nm. Light is green at this wavelength
What does addition of sodium dithionite do to haemoglobin?
What does oxygen binding grant Hb?
Altered e- resonance, so different absorption spectrum to other proteins
Hb with Fe3+ instead of fe2+, caused by oxidation.
This is what gives dried blood its brown colour.
What is the Beer-Lambert Law?
A = E x c x I
Absorbance = Extinction coefficient x concentration x path length.
Basically says that the absorbance of a solution is proportional to the concentration of the absorbing solute and to the distance travelled by light through the sample.
It can be used to determine Hb conc in a sample using spectrophotometry results
Which is more negatively charged. HbA or HbS?
HbA is more negative