Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Primary E12 mouse cortex)
Specification
Primary E12 mouse cortex
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Other product details
Dilution
1/250
Incubation time
24 hour(s) and 0 minute(s)
Secondary antibody
Secondary antibody
None used
Additional data
Additional Notes
Fix with 2% para for 15mins followed by 95% methanol for 30mins at RT.
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.
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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提交于 Feb 22 2007