Interpretation Of Results - helena BioSciences SAS-1 Immunofix Instrucciones De Uso

Stain
Destain
Destain
Dry
c) Manual
Follow the sequence listed for the SAS-2 staining unit, using a staining bath for the Stain, Destain
and Wash steps, and a Drying Oven with forced air at 60...70°C for the Dry steps.
18. At the end of the staining cycle, remove the gel from the staining chamber. The gel is now ready
for examination.

INTERPRETATION OF RESULTS

The majority of monoclonal proteins migrate in the cathodic, gamma region of the protein pattern,
but due to their abnormal nature, they may migrate anywhere within the globulin region on protein
electrophoresis. The monoclonal protein band on the immunofixation pattern will occupy the same
position and shape as the abnormal band on the serum protein pattern. The abnormal protein is
identified by the antiserum type it reacts with.
When low concentrations of abnormal protein are present, the abnormal band may appear as a band
within the normal polyclonal immunoglobulin. A band can also be seen within a polyclonal background
when there is a large polyclonal immunoglobulin presence also.
The publication 'Immunofixation for the Identification of Monoclonal Gammopathies' is available from
Helena BioSciences on request.
Urine Immunofixation:
Type of Proteinuria
Normal urine
Glomerular
Tubular
Overflow
LIMITATIONS
1. Antigen Excess
Antigen excess will occur if there is not a slight antibody excess or antigen / antibody equivalence
at the site of precipitation. Antigen excess in IFE is usually due to an excess of the immunoglobulin
in the patient sample. Antigen excess is characterised by prozoning (unstained areas in the centre
of the immunofixed protein band, with staining around the edges). A higher dilution of the sample
should be used in this event to optimise the immunoglobulin concentration.
2. Non-Specific Precipitation in All Immunoglobulin Lanes
Occasionally a completed IFE plate exhibits a precipitate band in the same position in every pattern
across the plate. This may result from:
a) IgM monoclonal immunoglobulins.
04:00
02:00
02:00
12:00
Bands Observed On Gel
Small albumin band
Albumin, alpha-1,
beta, gamma
alpha-1, alpha-2, beta
gamma or variable
63
6
Recirculate ON
Recirculate ON
Recirculate ON
Proteins Present
albumin
albumin, alpha-1
antitrypsin, transferrin,
gamma globulins
Retinol Binding Protein,
beta2-microglobulin,
alpha-2 microglobulin.
immunoglobulins,
free light chains