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
Divulgations
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 |
Tags
Citer Cet Article
Immunofluorescence Assay to Evaluate Effect of Target Proteins on Neurite Outgrowth. J. Vis. Exp. (Pending Publication), e21331, doi: (2023).