Immunofluorescence Assay to Evaluate Effect of Target Proteins on Neurite Outgrowth

Published: April 30, 2023

Abstract

Source: Chan, W., W., R. et al. An Enhanced Green Fluorescence Protein-based Assay for Studying Neurite Outgrowth in Primary Neurons. J. Vis. Exp. (2019).

In this video, we demonstrate an EGFP-based immunofluorescence assay to identify the efficacy of target proteins on the neurite outgrowth of primary cortical neurons. The primary neurons are transfected with a fluorescent protein, EGFP, and a target protein, and then an immunoassay is performed to investigate the effect of the target protein on neurite outgrowth.

Introduction

Açıklamalar

The authors have nothing to disclose.

Materials

18 mm Circle Cover Slips Thermo Scientific CB00180RA Sterilize before use
Cytochalasin D Invitrogen PHZ1063 Dissolved in DMSO
EndoFree Plasmid Maxi Kit QIAGEN 12362
Fluorescence Mounting Medium Dako S302380
Lipofectamine 2000 Transfection Reagent Invitrogen 11668019
Paraformaldehyde Sigma-Aldrich P6148
pEGFP-C1 Clontech #6084-1
pCI FE65 Please see references 8 and 15
PBS Tablets Gibco 18912014
Penicillin-Streptomycin Gibco 15140122
Poly-D-lysine hydrobromide Sigma-Aldrich P7280
Spatula Sigma-Aldrich S4147
Trypsin-EDTA (0.05%), phenol red Gibco 25300062

Etiketler

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Bu Makaleden Alıntı Yapın
Immunofluorescence Assay to Evaluate Effect of Target Proteins on Neurite Outgrowth. J. Vis. Exp. (Pending Publication), e21331, doi: (2023).

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