helena BioSciences SAS-1 Immunofix Instrucciones De Uso página 6

i i i i ) ) S S A A S S - - 1 1 P P l l u u s s u u s s e e r r s s : : dispense 400µL of REP Prep onto the heat sink. Place the gel onto the heat sink,
agarose side up, aligning the positive and negative sides with the corresponding electrode posts,
taking care to avoid air bubbles under the gel.
i i i i i i ) ) S S A A S S - - 3 3 u u s s e e r r s s : : place the alignment guide onto the pins and dispense 400µL of REP Prep onto the
centre of the chamber. Place the gel into the chamber agarose side up, using the guide, align the
positive and negative sides with the corresponding electrode posts, taking care to avoid air bubbles
under the gel.
3. Blot the surface of the gel with a blotter C, discard the blotter.
4. i i ) ) S S A A S S - - 1 1 u u s s e e r r s s : : attach the electrodes onto the top side of the electrode posts so that they are in
contact with the gel blocks.
i i i i ) ) S S A A S S - - 1 1 P P l l u u s s u u s s e e r r s s : : (as above). Place the cover over the gel and electrodes and press firmly for 5
seconds to ensure contact.
i i i i i i ) ) S S A A S S - - 3 3 u u s s e e r r s s : : attach the electrodes onto the the electrode posts so that they are in contact with
the gel blocks.
5. Place 2 applicator blade assemblies in position on the instrument, ( ( S S A A S S - - 3 3 u u s s e e r r s s : : s s l l o o t t A A a a n n d d 1 1 0 0 ) ) .
6. Perform the Immunofix electrophoresis:
i i ) ) S S A A S S - - 1 1 u u s s e e r r s s : : 80 volts, 20 mins
i i i i ) ) S S A A S S - - 1 1 P P l l u u s s u u s s e e r r s s : : Electrophoresis: 100 volts, 18 mins, 20°C
i i i i i i ) ) S S A A S S - - 3 3 u u s s e e r r s s : :
Step
Load Sample
Apply Sample
Electrophoresis
Apply antisera
Insert Combs
Blotter D
Dry
* Use Location 2
N N O O T T E E 1 1 : : For Serum immunofixation, 1 sample application is required. For Urine immunofixation,
10 sample applications are required. Remove the gel blocks prior to drying.
N N O O T T E E 2 2 : : Urine and serum samples can be run simultaneously on one gel. However, each
individual row must contain one type of sample only (eg. top row = urine, bottom row = serum).
If a combination of serum and urine are to be used on the same gel, place both blades onto the
SAS-1plus, and remove the 'serum' blade after the first load/application. Leave the 'urine' blade to
complete the other 9 load/applications.
7. Following electrophoresis, ( ( S S A A S S - - 1 1 P P l l u u s s u u s s e e r r s s : : remove the cover), remove the electrodes from the
surface of the gel. ( ( S S A A S S - - 3 3 u u s s e e r r s s : : remove the alignment guide). 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.
8. Apply 2 drops (or 50µL) of Protein Fixative (serum) or Total Antiserum (urine) into the hole of the
SP lane and 2 drops (or 50µL) of the appropriate antiserum into the hole of the immunoglobulin
lanes. Ensure that the fixative and antisera have completely filled the channels.
9. Incubate the gel. ( ( S S A A S S - - 1 1 u u s s e e r r s s : : incubate at 15...30°C).
Incubation Step 1: 8 mins, 37°C (incubate)
Incubation Step 2: 8 mins, 40°C (D blot)
Time (mm:ss)
00:30
00:30
17:00
10:00
02:00
05:00
08:00
Temperature (°C)
21
21
21
21
21
40
54
4
Voltage Other
Speed 1
Speed 1*
100