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Immunoelectrophoresis (IEP), Serum Protein Electrophoresis (SPE) & Immunofixation (IFX)
Immunoelectrophoresis (IEP), Serum Protein Electrophoresis (SPE) & Immunofixation (IFX)
Mixed antigens
Antiserum B
Well : antigen mixture (usually a serum sample)
Troughs : antibody/mixture of antibodies
Electrophoresis : to separate proteins in the sample
-antigens and antibodies migrate toward 1 another
precipitin arcs at the region of optimal concentration
4. Description
When serum applied to the agarose plate
subjected to normal zone electrophoresis
Channels that are filled with specific antiserum
Chemical
Separat methods
Electrophore
Electrophore
e sis
sis
Elemen Ultracentrifu
ge
A process of separating electrically
charged particles in solution Protein
by passing
Immunoelectrophor
an electric current through the solution
Electrophores
esis
is
assess the blood levels of the total amount of protein
specific types of proteins in the blood, and to
called immunoglobulins. establish the levels of other
types of proteins - albumin,
ordered if a SPE test has a
Electrophore
"spike," or rise, at the
alpha1 globulin, alpha2
globulin, beta-globulin &
immunoglobulin level sis Gamma globulin.
Albumin &
Globulin
SP
Serum protein electrophoresis uses an
electrical field to separate the proteins in the
blood serum into groups of similar size,
shape & charge.
25 min, 100V
Destain
Ge l E lectro phor esi sii
-ve charge conferred on molecules
gel is porous
molecules travel through gel according to
size when electrical current applied
smaller travel faster
Migration rate
-Charge of the molecule
-Size & shape of the molecule
-Voltage
-Support medium
-pH & ionic strength of the buffer
PRINCIPLE
Serum sample is applied closed to the cathode
All major serum protein carry a net –ve charge at pH 8.6
migrate toward the anode after current passed through
Serum protein is arranged into 5 bands
Albumin, α1 –globulin,α2-globulin,β-globulin,ɣ-globulin
toward the
Anode anode Catode
(+) (-)
Intrepretation of
result
Abnormal result
Continue
Abnormal result
WEAKNESSES Not be able to have the test or
effect the results