lecture 6 techniques in protein chemistry
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Lecture 6
Techniques inProtein
Chemistry
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UV-Visible spectrophotometer
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UV-Visible spectrophotometer
• It uses two light sources, a deuterium (D2)lamp for ultraviolet light and a tungsten ()lamp for visible light!
• "fter bouncing off a mirror (mirror #), the
light beam passes through a slit and hits adiffraction grating!
• $he grating can be rotated allowing for aspecific wavelength to be selected!
• "t an% specific orientation of the grating, onl%monochromatic (single wavelength)successfull% passes through a slit!
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Isoelectric 'ocusing
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"pplied ios%stems rocise +# $
rotein .e/uencer
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igh erformance 0i/uid
1hromatograph% (01)
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Staining of PVDF-bound standard proteins. .tandard proteins (indicated b%
arrows) used were human transferrin (34), bovine serum albumin (564), rabbitmuscle actin (+34), so% bean tr%psin inhibitor (2#4), and horse heart m%oglobin(#+4)! 0anes from left to right were #) io-7ad molecular weight mar8ers, 29:)2; pmol each standard protein, +93) #; pmol each standard protein, 59) 3 pmoleach standard protein, 69) 2 pmol each standard protein, and #;) blan8 lane!
"ll staining conditions were as described above! 1 is an abbreviation for
1oomassie rilliant lue