Cryosections from embryos collected at E12.5 were prepared and processed for staining as in Charoy et al., 2012 (link). For some experiments, chick embryo sections and open-book spinal cords were blocked in 6% BSA (A7906, Sigma) and 0.5% Triton (T9284, Sigma) diluted in PBS for 5 hr at room temperature. Sections were incubated overnight at room temperature with anti-PlxnA1 antibody (gift from Y. Yoshida), anti-Robo3 antibody (1/100, R and D, AF3076); anti-L1CAM antibody (1:100, A439 Abcam 123990), anti-NgCAM antibody (1:50, 8D9, DSHB), anti-BEN (1:50, BEN, DSHB) or an anti-PC2 antibody (1:100, 3533, Abcam) in 1% BSA diluted in PBS. Alexa 488, Alexa 555 (1/500, Invitrogen) and Fluoroprobe 546 (1/400) were used as secondary antibodies.
Fluoroprobe 546
Manufactured by Thermo Fisher Scientific
The Fluoroprobe 546 is a fluorescence-based analytical instrument. It is designed to detect and quantify fluorescent substances in liquid samples. The Fluoroprobe 546 utilizes excitation and emission wavelengths to identify and measure the concentration of target analytes.
Automatically generated - may contain errors
2 protocols using fluoroprobe 546
1
Immunostaining of Embryonic Tissue Sections
2
Cryosectioning and Immunostaining of Embryonic Tissue
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Cryosections from embryos collected at E12.5 were prepared and processed for staining as in Charoy et al, (2012) . For some experiments, chick embryos sections and open-book spinal cords were blocked in 6% BSA (A7906, Sigma) and 0.5% Triton (T9284, Sigma) diluted in PBS for 5 hours at room temperature. Sections were incubated overnight at room temperature with anti-PlxnA1 antibody (gift from Y. Yoshida), anti-Robo3 antibody ((1/100, R&D, AF3076); anti-L1CAM antibody (1:100, A439 Abcam 123990), anti-NgCAM antibody (1:50, 8D9, DSHB), anti-BEN (1:50, BEN, DSHB) or an anti-PC2 antibody (1:100, 3533, Abcam) in 1% BSA diluted in PBS.
Alexa 488, Alexa 555 (1/500, Invitrogen) and Fluoroprobe 546 (1/400) were used as secondary antibodies.
Alexa 488, Alexa 555 (1/500, Invitrogen) and Fluoroprobe 546 (1/400) were used as secondary antibodies.
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