Immunocytochemistry for BrdU labeling of proliferative LSCs was performed as in our previous study [25 (link)]. Deparaffinized mouse eye sections were blocked with 10% goat serum and 5% BSA in PBS containing 0.1% Tween-20 for 1 h at RT. The eye sections were double-stained with antibodies to rabbit polyclonal anti-ΔNp63α antibody (1:100; BioLegend) and monoclonal anti-mouse Hes1 antibody (1:100; ab87395; Abcam) or monoclonal anti-mouse Smad7 antibody (1:100; sc-365846; Santa Cruz Biotechnology) at 37 °C for 2 h. The sections were then incubated with rhodamine-conjugated donkey anti-mouse IgG antibody and fluorescein isothiocyanate (FITC)-conjugated donkey anti-rabbit IgG antibody (1:500; Santa Cruz Biotechnology) for 1 h at RT. Nuclei were monitored by counterstaining with Hoechst 33,258 for 7 min. Images were captured using a Zeiss epifluorescence microscope(Oberkochen, Germany) with a charge-coupled device (CCD) camera and photographs taken using the Axiovert software.
Ab87395
Manufactured by Abcam
Ab87395 is a lab equipment product offered by Abcam. It is a high-quality tool designed for laboratory use. The core function of this product is to assist researchers and scientists in their work, but a detailed description while maintaining an unbiased and factual approach is not available.
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2 protocols using ab87395
1
Immunocytochemical Profiling of Proliferative LSCs
2
Immunohistochemical Analysis of HES1 Expression
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Formalin-fixed paraffin-embedded specimens were processed for fine sections (5 μm) and were mounted on slides coated with poly-l -lysine (Sigma, St Louis, MO), followed by conventional hematoxylin and eosin staining for histologic assessment and immunohistochemistry (IHC). Citrate buffer (pH 6.0) was used for antigen retrieval treatment in the microwave and incubated overnight at 4°C with the primary rabbit polyclonal antibody of HES1 (ab87395; Abcam, Cambridge, United Kingdom) at a dilution of 1:250 respectively. An Envision System peroxidase kit (DAKO, Carpinteria, CA) was used for staining. Detailed methodology has been described earlier.19 (link)
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