Hoefer PR625 Immunoblot - Manual (Page 5)

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p3
Instructions
The following detail the steps for probing a
membrane using the ImmunoBlot.
Select and block membrane
1
Select a membrane of choice (nitrocellulose,
Hybond ECL
™ ; PVDF, Hybond P; low fluorescent PVDF,
Hybond LFP) for the assay and cut to:
Length: 15 cm
Width: Cut to accommodate channels to be used
Maximum width for all channels: 15 cm
2
Block membranes with excess non-specific protein. 1
3
Note: If blocked membranes have been stored in a
protein-free solution, re-block them for 1 to 2 minutes
in a tray of blocking solution.
Load membrane
1
Remove the top acrylic plate of the ImmunoBlot
and turn it over.
2
With the antigen-bearing face of the membrane facing
the ImmunoBlot channels, position the membrane so
it covers all the channels.
2
3
Place a new, dry sealing pad (supplied with unit) over
the membrane.
4
Set the bottom plate of the unit on the inverted top
plate so the alignment pins fall in place.
5
Ensure there is no gap between the top and
bottom plates.
1 See Appendix A, Note 1,
Blocking the membrane.
2 See Appendix A, Note 2,
Aligning the membrane.