Hoefer PR625 Immunoblot - Manual (Page 7)

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p5
Remove primary antibody solutions
and wash the blot
Use the washing manifold to remove the
antibody solution and wash the blot.
1
Position the 2 identical parts of the washing manifold
in the slots on either side of the top plate and press
firmly until the O-rings are seated in the slots.
5
2
To aspirate all samples simultaneously, first connect
pieces of the tubing supplied with the unit to both
parts of the manifold using a luer fitting. Now,
connect the open end of the tubing from the manifold
piece to a trap connected to a vacuum source.
Then place the open end of the tubing from the
second manifold piece into a beaker containing
wash buffer.
When you start the vacuum antibody solutions in the
channels are removed and the wash buffer is drawn
in from the other side. Please refer to your Western
detection protocol for wash instructions. For Western
detection we recommend at least 2 quick washes
in buffer followed by two longer washes in buffer
(5 min while rocking is sufficient) before introducing
the secondary antibodies.
5 See Appendix A, Note 5, Fitting
the O-rings into the slots.