Millipore SNAP i.d User Manual

Before using the SNAP i.d. Protein Detection System, please read the User Guide completely.
1. Open the blot holder lid, taking
care not to damage the inner
white surface.
2. Thoroughly wet the white
surface of the blot holder with
Milli-Q ® water.
NOTE: If using only one
well of a double or triple
well blot holder, the unused
well(s) must also be wet.
3. Place the pre-wet blot in the
center of the blot holder with
the protein side down. The blot
membrane should not exceed
size specified in the User Guide.
4. Roll blot membrane gently to
remove air bubbles.
5. Place the spacer (wetting not
necessary) on top of the blot
membrane and roll again to
ensure contact of spacer with
blot membrane.
6. Close the blot holder lid.
Squeeze firmly at base of tab
7.
area to secure lid.
00103870 Rev. A, 01/08
OVERVIEW OF PROCEDURE
8. Open lid of system and place
blot holder in chamber, aligning
blot holder tabs with notches of
chamber. Close and latch lid.
9. Add volume of blocking solution
as indicated under OPTIMIZA-
TION GUIDELINES on reverse
side. Using knobs on the system,
apply vacuum until well(s) are
completely empty.
TURN VACUUM OFF.
10. Add volume of primary anti-
body as indicated under
OPTIMIZATION GUIDE-
LINES on reverse side.
Antibody solution must evenly
cover entire blot holder
surface.
Incubate for 10 minutes at
room temperature. Solution
will be absorbed into the blot
holder and surface may ap-
pear dry. Apply vacuum.
IMPORTANT: Do not ap-
ply vacuum until after the
10-minute incubation.
11. With vacuum running
continuously, wash 3 times
with wash buffer. See
OPTIMIZATION GUIDE-
LINES on reverse side for
volumes.
TURN VACUUM OFF.
12. Apply appropriate volume of secondary antibody (see
GUIDELINES on reverse side) evenly across the blot holder surface.
Incubate for 10 minutes at room temperature. Again, solution will be ab-
sorbed into the blot holder and surface may appear dry. Apply vacuum.
IMPORTANT: Do not apply vacuum until after the 10-minute
incubation.
13. With vacuum on, wash 3 times with buffer. See
GUIDELINES on reverse side for volumes.
TURN VACUUM OFF.
14. Remove blot and incubate with the
appropriate detection reagent such
®
as Immobilon HRP, or, if using Milli-
pore fluorescently labeled antibod-
ies, visualize.
Millipore, Milli-Q and Immobilon are registered trademarks of
Millipore Corporation.
SNAP i.d. and the M mark are trademarks of Millipore Corporation.
Tween is a registered trademark of ICI Americas Inc.
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OPTIMIZATION
OPTIMIZATION