Interpretation Of Results - helena BioSciences SAS-3 Immunofix Instrucciones De Uso

minutes for the excess antisera to be absorbed, then remove the blotter combs and the template
from the gel surface.
12. Place a blotter D (smooth side down) onto the surface of the gel and replace the antiserum
template to hold the blotter flat. Blot the gel.
13. Remove the blotter D and dry the gel.
NOTE: Immediately after use, clean the antisera template with a mild, biocidal detergent. If
possible, scrub the bottom of the template with a toothbrush or small test tube brush. Do not let
the antisera dry on the template. Build up of antisera on the surface of the template will result in
the formation of bubbles during the antisera application step. Dry the antisera template throughly.
Water left in the holes will hinder the application of antisera for the next use. Store the template
upside down to increase the air circulation and thus the drying potential of the applicator.
14. Attach the gel to the staining chamber holder.
15. Select the IFE test program on the staining unit and, following the prompts, Wash, Stain and Dry
the gel.
16. 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 excessis 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.
Bands Observed On Gel
Small albumin band
Albumin, alpha-1,
beta, gamma
alpha-1, alpha-2, beta
gamma or variable
5
SAS-3 IMMUNOFIX
Proteins Present
albumin
albumin, alpha-1
antitrypsin, transferrin,
gamma globulins
Retinol Binding Protein,
beta2-microglobulin,
alpha-2 microglobulin
immunoglobulins,
free light chains
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