ELECTRIPHOREISIS AND ELECTROCHEMISTRY Flashcards
→ A molecule that contains both acidic and basic groups
Ampholyte
The rate of migration of a charged solute in an electric
field, expressed per unit field strength.
● Electrophoretic Mobility
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.
Endosmosis
→ The migration of charged macromolecules.
Electrophoretogram
The migration of small charge ions.
Iontophoresis
The migration of charged macromolecules.
Zone Electrophoresis
is acetylated to form cellulose acetate by treating it
with acetic anhydride
Cellulose acetate
Different fractions of lipoproteins
based on their density
High-density lipoproteins (HDL)
▪ Low-density lipoproteins (LDL)
▪ Intermediate-density lipoproteins
(IDL)
▪ Very low-density lipoproteins (VLDL)
negatively charged
Cathode
positively charged
Anode
Versatile and powerful analytical technique capable of
separating and analyzing a diverse range of ionized analytes.
ELECTROPHORESIS
Macromolecules of interest:
Proteins in serum
→ Urine
→ Cerebrospinal fluid (CSF)
→ Erythrocytes and tissue and,
→ Other biologic body fluids
Separates by electrical charge
● A highly purified uncharged polysaccharide derived from agar
● Neutral; therefore, doesn’t produce electroendosmosis
Agarose gel
CONVENTIONAL ELECTROPHORESIS (4)
→ Separation
→ Staining
→ Detection
→ Quantification
Referred to as PAGE
Involves separation of protein on the basis of charge and
molecular size
● Layers of gel with different pore sizes are used
Polyacrylamide gel
Used to separate macromolecules on the basis of both
surface charge and molecular charge
Starch
Power supplies operating at either constant current or
constant voltage are available commercially.
DRIVING FORCE (ELECTRICAL POWER)
is a molecule whose net charge can be either
positive or negative
AMPHOLYTE
Serves as a multifunctional component in the electrophoretic
process
BUFFER
Serum protein separation,
Poor resolution, weak buffer
Barbitone buffer –
(around 8.0 pH)
Enzyme separation,
Low buffering capacity, high
conductivity
Phosphate buffer
– (around 7.0 pH)
Nucleic acid separation
Good resolution, high buffering
capacity, low conductivity
Tris – borate –
EDTA buffer
(TBE) – (pH
around 8.0)
Nucleic acid separation
High resolution, high buffering
capacity, low conductivity
Tris – acetate –
EDTA buffer
(TAE) – (pH
around 8.0)
Protein separation
High buffering capacity, low
conductivity
Tris – glycine
buffer – (pH more
than 8.0)
Based on refractivity (the ability of the
substance to bend light)
REFRACTOMETRY
Involves the measurement of electrical signals associated
with chemical system that are incorporated into an
electrochemical cell
ELECTROCHEMISTRY
Converts chemical energy into electrical energy
● The redox reaction is spontaneous and is responsible for the
production of electrical energy
GALVANIC CELL