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Permeabilise with 1N HCl on ice for 10mins followed by 2N HCl for 10mins at RT.
Immediately after acid washes, incubate with 0.1M sodium borate for 12mins at RT. Block for 1 hour with 5%NGS with 0.1% Triton. Incubate overnight with primary antibody +0.1% TX + 10% serum.
To visualise this biotinylated primary, streptavidin conjugated to Alexa488 fluorochrome was used at 1:500 from Molecular Probes (Invitrogen) for 1 hour. Image shows BrdU staining, with nuclei counter stained with DAPI.
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Submitted Feb 22 2007