Antibodies Used for Neuronal Immunostaining

Antibodies used for immunostaining included: mouse anti-nc82 (1:50, Developmental Studies Hybridoma Bank), rat-anti-Chinmo (1:500; Wu et al., 2012 (link)), rabbit anti-Br-Z3 (1:250; this study), rabbit anti-GFP (1:1000; Invitrogen), chicken anti-GFP (1:500; Rockland Immunochemicals), rabbit anti-dsRed (1:500; Clontech), mouse anti-mCherry (1:500; Clontech), rabbit anti-β-gal (1:500; MP Biomedicals), chicken anti-β-gal (1:500; Abcam). Most primary antibodies were preabsorbed against fixed embryos. Secondary goat antibodies were conjugated to Alexa Fluor 488, 568 or 633 (Molecular Probes). Slides were mounted in Vectashield (Vector Labs) or dehydrated through an ethanol series and mounted in DPX. Images were collected on a Leica SP5 (Light Microscopy Imaging Center, Indiana University). Confocal stacks were merged using Leica LSM software. Samples that were directly compared were prepared under identical conditions and imaged in parallel.

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Wu Y.C., Chawla G, & Sokol N. (2020). let-7-Complex MicroRNAs Regulate Broad-Z3, Which Together with Chinmo Maintains Adult Lineage Neurons in an Immature State. G3: Genes|Genomes|Genetics, 10(4), 1393-1401.

Publication 2020

mouse anti-nc82 rabbit anti-GFP chicken anti-GFP rabbit anti-dsRed mouse anti-mCherry rabbit anti-β-gal chicken anti-β-gal Vectashield

Alexa fluor 488 Antibodies Chicken Dehydrated ethanol Embryos Goat Hybridoma Light microscopy Molecular probes Mouse Rabbit Vector

Corresponding Organization :

Other organizations : Indiana University Bloomington

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Variable analysis

independent variables

Antibodies used for immunostaining

dependent variables

Immunostaining patterns

control variables

Identical conditions and parallel imaging for samples that were directly compared

controls

Most primary antibodies were preabsorbed against fixed embryos (positive control)
No negative controls were explicitly mentioned

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