Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BrdU antibody (Biotin)

Excellent
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Mouse tumour)
Specification
Mouse tumour
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
Perkin-Elmer TNB Blocking Reagent as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: 25°C

Other product details

Dilution
1/500
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: TNB

Secondary antibody

Secondary antibody
None used

Additional data

Additional Notes
Animals given 1.5mg BrdU pulse at least 2 hours prior to transcardial perfusion with saline followed by 4% paraformaldehyde. Sections were cryoprotected, frozen and cut at 15µm on cryostat (NOT PARAFFIN EMBEDDED). Amplified GFP in tumour cells with anti-GFP antibodies followed by Alexa488 conjugated secondary antibodies prior to BrdU protocol as HCL destroys GFP fluorescence. Used BrdU protocol as published by abcam with the modification of a single 30min incubation at 37C in HCL rather than at different temperatures. For development of fluorescence we used Perkin-Elmer tyramide signal amplification for biotin followed by Alexafluor546 conjugated streptavidin. This product is extremely clean. Beautiful staining. Positive control was RMS/SVZ.

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Submitted Feb 22 2008

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