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Cell Signaling & Neuroscience
Transfecting Cultured Neurons
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Selected Protocol from ""Live Cell Imaging", D. Spector & R. Goldman.
Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid
D. Spector & R. Goldman
ABSTRACT
This protocol describes two transfection methods for expressing GFP-tagged actin in primary neurons. The lipid reagent DOTAP (Roche Diagnostics) method produces actin-GFP-expressing hippocampal neurons that survive well during long periods in culture. The calcium phosphate method can be used to transfect neurons that have already been growing on coverslips in vitro. Transfected cells suitable for imaging can be obtained in cultures up to 15 days in vitro. One to two percent transfected cells is a typical result. A disadvantage of the calcium phosphate method is that hippocampal neurons become "fragile" after treatment.
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Products Available for this Protocol
| Protocol Material Description |
Product # |
Product Name |
Add to Cart |
| Buffers, Solutions, and Reagents |
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| CaCl2 |
C2661 |
Calcium chloride, plant cell culture tested, anhydrous |
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| NaCl |
S3014 |
Sodium chloride, for molecular biology |
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| KCl |
P9541 |
Potassium chloride, for molecular biology |
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| Na2HPO4 |
71649 |
Sodium phosphate dibasic dodecahydrate |
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| BES-buffered saline (BBS), 2x |
14280 |
BES buffered saline, for molecular biology, 2x concentrate |
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| BES |
B4554 |
BES, Biotechnology Performance Certified, cell culture tested |
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| NaCl2 |
S3014 |
Sodium chloride, for molecular biology |
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| Glucose |
G7528 |
D-(+)-Glucose, SigmaUltra |
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| Hank's balanced salt solution (HBSS); Ca++-Mg++-free |
H4385 |
Hanks' Balanced Salt solution, 10 ×, Modified, without calcium, magnesium or sodium bicarbonate |
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Product Association Disclaimer: The Sigma-Aldrich products listed for this specific protocol were selected either to match or to supplement the products listed within the actual protocol. The products/reagents from Sigma-Aldrich have been qualified for usage, but may not have been validated for this specific application. Please refer to the detailed product description on the usage of specific products of interest.
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