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.
Apply appropriate volume of secondary antibody (see
12.
OPTIMIZATION
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.
With vacuum on, wash 3 times with buffer. See
13.
OPTIMIZATION
GUIDELINES
on reverse side for volumes.
TURN VACUUM OFF.
Open lid of system and place
8.
blot holder in chamber, aligning
blot holder tabs with notches of
chamber. Close and latch lid.
Add volume of blocking solution
9.
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.
Add volume of primary anti-
10.
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.
+
+
With vacuum running
11.
continuously, wash 3 times
with wash buffer. See
OPTIMIZATION GUIDE-
LINES
on reverse side for
volumes.
TURN VACUUM OFF
.
3X
Remove blot and incubate with the
14.
appropriate detection reagent such
as Immobilon
®
HRP, or, if using Milli-
pore fluorescently labeled antibod
-
ies, visualize.
Squeeze firmly at base of tab
7.
area to secure lid.
Close the blot holder lid.
6.
Thoroughly wet the white
2.
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.
Place the spacer (wetting not
5.
necessary) on top of the blot
membrane and roll again to
ensure contact of spacer with
blot membrane.
Roll blot membrane gently to
4.
remove air bubbles.
Open the blot holder lid, taking
1.
care not to damage the inner
white surface.
OVERVIEW OF PROCEDURE
Before using the SNAP i.d. Protein Detection System, please read the User Guide completely.
Place the pre-wet blot in the
3.
center of the blot holder with
the protein side down. The blot
membrane should not exceed
size specified in the User Guide.
00103870 Rev. A, 01/08
AVID QS 021108.indd 1
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