helena BioSciences SAS-3 Immunofix Instrucciones De Uso página 6

NOTE 1: For Serum immunofixation, 1 sample application is required. For Urine immunofixation,
10 sample applications are required.
NOTE 2: Urine and serum samples can be run simultaneously on one gel. However, each
individual row must contain one type of sample only (eg. rows 2 and 8 = urine, row 14 = serum).
If a combination of serum and urine are to be used on the same gel, ensure that the blades for
urine samples go onto the SAS-3 for the first 9 load/applications. Add applicator blade for serum
sample for the final (10th) load/application only.
Step
Stainer
Wash
Wash
Stain
Destain
Destain
Dry
2. Place the correct number of applicator blade assemblies in position on the instrument:
Number of Samples
9
6
3
3. i) SPIFE Combo users: Pipette 17µl of the sample into the appropriate well of the sample base.
Protect the samples from evaporation.
ii) SPIFE 2000 / 3000 / SAS-3 users: Pipette 35µl of the sample into the appropriate disposable
sample cups. Protect the samples from evaporation.
4. Remove the gel from the packaging and discard the overlay.
5. Dispense 2ml of REP Prep onto the left side of the chamber and carefully place the gel into the
chamber, aligning the holes in the gelbond with the pins in the chamber and avoiding bubbles under
the gel. Attach the electrodes onto the outside of the electrode posts so that they are in contact
with the gel blocks.
6. Blot the surface of the gel with a Blotter C, discard the blotter.
7. Select the IFE test program and, following the prompts, apply the samples and perform the
electrophoresis.
8. Following electrophoresis, remove the electrodes and remove both gel blocks using the Gel Block
Remover. Position the antiserum application template onto the gel surface. NOTE: The milled
antisera channels should be aligned centrally over the printed box on the gel in which the samples
are applied.
9. Apply 2 drops (or 60µL) of Protein Fixative (serum) or Total Antiserum (urine) into the top hole
of the SP lane and 2 drops (or 60µL) of the appropriate antiserum into the top hole of the
immunoglobulin lanes. Ensure that the fixative and antisera have completely filled the channels.
10. Incubate the gel.
11. Following incubation, place a blotter comb into the top holes of the antiserum template. Allow 2
Time (mm:ss) Temperature (°C)
00:03
10:00
04:00
02:00
02:00
12:00 63
SPIFE Combo
Applicator Position
4, 11, 18
4, 11
4
4
Other
Recirculate ON
Recirculate ON
Recirculate ON
Recirculate ON
Recirculate ON
SPIFE 2000/3000 and SAS-3
2, 8, 14
2, 8
2