Immunochemistry Analysis of Larval Brains

Larvae of desired genotype were dissected at 96 h ALH and brains were fixed for 15 min in 3.7% formaldehyde in PBS with 0.1% Triton-X, and later processed for immunochemistry analysis. The following antibodies were used: mouse anti-flag (Sigma), 1/2000; rabbit anti-Mira^{70 (link)} (generated in our lab), 1/1000; guinea-pig anti-Dpn (generated in our lab), 1/1000; rabbit anti-Pon (generated in our lab), 1/2000; rabbit anti-Numb, 1/1000; rabbit anti-aPKCζ C20 (Santa Cruz Biotechnologies, SG-216-G), 1/1000; chicken anti-GFP (Abcam, ab13970), 1/5000; anti-NICD (DSHB, C17.9C6), 1/50; anti-NECD (DSHB, C458.2H), 1/50; and anti-Ase (generated in our lab), 1/1000. Secondary antibodies were conjugated to Alexa Fluor 488, Alexa Fluor 555, or Alexa Fluor 633 (Molecular Probes), and used at 1/500, 1/1000, and 1/250, respectively. TO-PRO-3 (Invitrogen) was used at 1/5000 for DNA staining and samples were mounted in Vectashield (Vector Laboratories). Images were obtained using Leica SP8 upright microscope and processed in Adobe Photoshop CS6 and Adobe Illustrator CS6.

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Shan Z., Tu Y., Yang Y., Liu Z., Zeng M., Xu H., Long J., Zhang M., Cai Y, & Wen W. (2018). Basal condensation of Numb and Pon complex via phase transition during Drosophila neuroblast asymmetric division. Nature Communications, 9, 737.

Publication 2018

mouse anti-flag rabbit anti-aPKCζ C20 chicken anti-GFP Alexa Fluor 488 Alexa Fluor 555 Alexa Fluor 633 Vectashield Illustrator CS6

Alexa fluor 488 Alexa fluor 555 Antibodies Brains Chicken Formaldehyde Genotype Guinea pig Larvae Microscope Molecular probes Mouse Rabbit Vector

Corresponding Organization :

Other organizations : Shanghai Medical College of Fudan University, Huashan Hospital, Temasek Life Sciences Laboratory, National University of Singapore, Hong Kong University of Science and Technology, University of Hong Kong, Nankai University

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