week 6 Flashcards
(14 cards)
SDS (sodium dodecylsulfate)
used before and during gel electrophoresis part of SDS page
SDS (sodium dodecylsulfate) is … charged
negatively.
SDS elmininates the effect caused by differences in proteins’: shape, size, charge, mass
shape
true or false: SDS causes all proteins to have the same charge (negative)
kinda true? they are all negatively charged because of SDS, but they really have the same charge:mass RATIO. So they’re all negative, but bigger proteins will be more negative
… is the sole determinant of the migration rate
mass
what is beta-mercaptoethanol and what does it do?
it’s a reducing agent that’s including in the loading dye. it breaks di-sulfide bonds to make sure protein fully unfolds
why aren’t we boiling the proteins after breaking their bonds?
this is usually done by Smo is a very delicate protein
the proteins should run through the gel from neg/pos to neg/pos
neg to pos
western blotting
electrophoretic transfer of proteins from gel to a membrane (either nitrocellulose or nylon)
what can we use to block the nonspecific binding sites on the membrane (aka proteins on membrane that are not the antigen) that we transfer proteins onto after SDS PAGE?
milk, bovine serum, albumin, gelatin
what is iBlot used for
run proteins from gel to the membrane. current also flows through membrane from cathode (-) to anode (+)
different ways to detect proteins
stains, radioactivity, chromogenic, chemiluminescence
true or false: refractive index of oil is = to that of air.
false. refractive index of oil is = to that of GLASS. therefore, when light rays leave the glass slide and go through oil, it won’t bend. so then more rays can reach the objective
caspases are mostly in…
cytosol
