(a) Thaw protein ladder on ice. Mix well and aliquot the desired amount of the protein ladder (10 µl for mini-gels and 20 µl for full length gels) into a separate tube.
(b) Heat the ladder to 95-100ºC for 2 minutes.
(c) After a quick microcentrifuge spin, load directly onto gel. To ensure uniform mobility, load an equal volume of 1X Reducing SDS Loading Buffer into any unused wells.
(d) Peform Chemiluminescent detection according to manufacturer's instructions except, at the stage of secondary antibody incubation with the blot, concurrently add the anti-biotin HRP-linked antibody (provided) at a dilution of 1:1000. Incubate for 1 hour at room temperature with gentle agitation.
(e) Proceed as usual with the detection procedure
Biotinylated Protein Ladder: Store at -20°C.
Anti-biotin HRP-linked Antibody: Store at -20°C.
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