Fluorescence immunohistochemistry was performed as described previously.7 (link) Primary antibodies were rabbit anti-human Dishevelled-1 polyclonal antibody (PAJ540Hu01; Cloud-Clone, Houston, TX, USA), rabbit anti-human Dishevelled-3 polyclonal antibody (PAL453Hu01; Cloud-Clone), rabbit anti-human Frizzled3 polyclonal antibody (CSB-PA882067ESR2HU; Cusabio, Houston, TX, USA), rabbit anti-human Frizzled6 polyclonal antibody (G260; Assay Biotech, Fremont, CA, USA), mouse anti-human Prickle1 monoclonal antibody (sc-393034; Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-human Prickle2 polyclonal antibody (CSB-PA773783ESR1HU; Cusabio), mouse anti-human Vangl1 monoclonal antibody (sc-166844; Santa Cruz Biotechnology), and mouse anti-human Vangl2 monoclonal antibody (sc-515187; Santa Cruz Biotechnology). The antibodies were used at dilutions of 1:50.
Csb pa882067esr2hu
Manufactured by Cusabio
Sourced in United States
The CSB-PA882067ESR2HU is a laboratory equipment product. It serves as a core function related to scientific research and analysis, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation.
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2 protocols using csb pa882067esr2hu
1
Fluorescence Immunohistochemistry of Planar Cell Polarity Proteins
2
Immunofluorescence Imaging of Frizzled3 Receptor
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The specimens were fixed with 4% paraformaldehyde in Phosphate-Buffered Saline (PBS) at 4 °C overnight. The fixed samples were washed with PBS with 0.3% Triton X-100 (PBST), treated with 1.5% normal goat serum in PBST for 1 h, and incubated with rabbit anti-human Frizzled3 polyclonal antibody (CSB-PA882067ESR2HU; Cusabio) at 4 °C overnight. The primary antibody was used at a dilution of 1:50 in PBST containing 0.5% Bovine Serum Albumin (BSA). After a brief rinse with PBST, the samples were reacted at room temperature for 2 h with a secondary antibody, Alexa Fluor 488-conjugated goat anti-rabbit IgG (Invitrogen), diluted 1:1000 in PBST containing 0.5% BSA. After washing with PBS, the samples were placed with the apical surface downward in a thin-glass-bottomed Petri dish filled with PBS, gently pressed onto the bottom by metal weights, and examined under a Carl Zeiss LSM880 confocal laser-scanning microscope (Carl Zeiss).
The light source, an argon laser, was coupled to an Axio Observer7 inverted microscope (Carl Zeiss). Excitation wavelength was 488 nm. Laser power was reduced to 1.0% by a neutral density filter. An objective was Plan-Apochromat 40× with oil immersion. Emitted fluorescence was detected by a photomultiplier and displayed as a 512 × 512 pixel resolution image using Carl Zeiss ZEN 2 software.
The light source, an argon laser, was coupled to an Axio Observer7 inverted microscope (Carl Zeiss). Excitation wavelength was 488 nm. Laser power was reduced to 1.0% by a neutral density filter. An objective was Plan-Apochromat 40× with oil immersion. Emitted fluorescence was detected by a photomultiplier and displayed as a 512 × 512 pixel resolution image using Carl Zeiss ZEN 2 software.
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