ELECTRO Flashcards
(19 cards)
In electrophoresis, heat is produced when
current flows through a medium that has
resistance, resulting in an increase in
thermal agitation of the dissolved solute
(ions) and leading to a decrease in
resistance and an increase in current.
Driving force (electrical power)
It is negatively charged
Cathode
Positively charged
Anode
is a molecule whose net
charge can be either positive or negative
o If buffer is more acidic than the pI becomes
positively charged and migrates toward the
cathode
Amplolyte
It measures the absorbance of stain -
concentration of the dye and protein
fraction.
o It scans and quantitate electrophoretic
pattern.
Densitometry
the ability of the substance to bend light) when the light passes from one medium into another, the path of the light beam changes direction at the boundary surface if its speed in the second medium is different from that in the first
Refractometry
the ratio of the two speed of light
Refractive index
o Similar to PAGE
o Not widely used because of technical
difficulty in preparing the gel
Starch
layers of gel with different pore sizes are
used
o separates serum proteins into 20 or more fractions rather than the usual five fractions separated by cellulose acetate or agarose
o Best used for studying isoforms of individual proteins
Polyacrylamide gel
Preferential movement of water in one
direction through electrophoresis medium due to selective binding of one type of charge on the surface of the medium.
Endosmsis
type of electrophoresis usually used for
migration of macromolecules in a porous
support medium
Zone electrophoresis
type of electrophoresis limited to migration of small ions
Iontophoresis
The rate of migration of a charged solute in an electric field, expressed per unit field
strength.
Electrophoretic mobility
It acetylated to form cellulose acetate by treating it with acetic anhydride
o a dry, brittle film composed of about 80% air space; becomes pliable once air spaces fill with electrolytes
o Homogenous medium with uniform pore
size and does not absorb the protein
o Advantageous for separating proteins with
similar MW
Cellulose acetate
Highly purified uncharged polysaccharide
derived from agar
1
TRANS:
o Require small amounts of sample (approx. 2
mL)
o Does NOT produce electroendosmosis
o Does not bind protein and, therefore,
migration is not affected
Agarose gel
molecule that contains both acidic and
basic group
Ampholyte
- The movement of electrically charged
compounds in a medium resulting to their
separations based on their electrical
charges when an electric current is applied
Electrophoresis
Factors affecting rate of migration
Net electric charge of molecule
Size and shape of the molecule
Strength of electric field
Properties of Supporting Medium
Temperature of operation
