Serum Protein Electrophoresis (SPE)

Routine SPE uses a buffer with a pH of ______.

8.6

What are two common gels used in SPE?

Agarose and polyacrylamide

Generally, during SPE proteins move from ______ to ______.

cathode (-), anode (+)

Isoelectric point (pI) is the pH of the buffer at which the protein molecule is ______.

neutral

Each protein has its own ______.

pI

If buffer = pI, proteins will not ______.

migrate

If pH(buffer) > pI, protein will move toward the ______.

anode (protein will be -)

If pH(buffer) < pI, protein will move toward the ______.

cathode (protein will be +)

What parameters affect ZONE electrophoresis?

- pH

- Composition: TRIS or TEA

- Concentration and ionic strength

- Gel or paper support

Capillary SPE relies on direct spectrophotometric detection of proteins at ______nm.

200-225

Capillary SPE does not include ______.

staining

During capillary SPE, proteins migrate through the detector according to their ______.

electrophoretic mobility

Capillary SPE is often used for ______.

hemoglobin

Capillary SPE is trickier when used on ______.

urine

In capillary SPE, the proteins migrate from ______ to ______.

anode (+), cathode (-)

Opposite ZONE protein electrophoresis

The buffer of capillary SPE is ______.

acidic (~4)

Immunofixation electrophoresis identifies specific ______.

immunoglobulins (monoclonal spike)

Immunofixation electrophoresis is a ______ step procedure.

2

Immunofixation electrophoresis step 1

Electrophoresis

Immunofixation electrophoresis step 2

Overlay antisera, then stain

Immunofixation results

What do these results indicate?

IgG is present with Kappa light chains

In Zone protein electrophoresis, proteins migrate according to ______ ratio.

mass:charge

Zone protein electrophoresis staining ______ is proportional to the amount of protein.

intesity

Zone protein electrophoresis moves from ______ to ______.

cathode (-), anode (+)

Zone protein electrophoresis produces zones of proteins that are heterogenous and ______ one another.

separated from

Order of globulin bands

From darkest to lightest

Albumin - a1 - a2 - B1 - B2 - y (gamma)

a1 components

a1-acid glycoprotein, a1-antitrypsin, a1-fetoprotein

a2 components

Haptoglobin, a2 macroglobulin, ceruloplasmin

B components

Transferrin, B2-microglobulin, fibrinogen

N/A

skip

y (gamma) components

Igs, CRP

Fraction Identification on a graph

From left to right

Albumin - a1 - a2 - B1 - B2 - -y (gamma)

CSF protein electrophoresis is common for ______ patients.

multiple sclerosis

______ bands present in CSF protein electrophoresis are likely indicative of multiple sclerosis.

Oligoclonal

Oligoclonal bands

Urine protein electrophoresis specimens need to be ______ due to low protein levels.

concentrated (same as CSF)

With Bence Jones proteins in urine PE, ______ are found loose in the urine of Multiple Myeloma patients.

light chains

Cryoglobulins are Igs in the serum that ______ at low temperatures.

precipitate