In-house made nucleofection buffer for efficient and cost effective transfection of RAW 264.7 macrophages
| Autor: | Bandish Kapadia, Kishore V.L. Parsa, Neeraja P. Alamuru-Yellapragada |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cost-Benefit Analysis Biophysics Ficoll India Nucleofection Buffers Biology Protein Engineering Transfection Biochemistry Buffer (optical fiber) Effective solution Mice 03 medical and health sciences Animals Molecular Biology RAW 264.7 Cells Electroporation Cell Biology Poloxamer Molecular biology Recombinant Proteins Cell biology 030104 developmental biology Batch Cell Culture Techniques Culture Media Conditioned |
| Zdroj: | Biochemical and Biophysical Research Communications. 487:247-254 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2017.04.043 |
| Popis: | Electroporation is the most widely employed method of gene transfer into macrophages which are hard to transfect. RAW 264.7 is a widely used cell line for studying macrophage responses. Electroporation of RAW 264.7 cells with commercial reagents although very efficient is expensive necessitating the development of cost effective alternatives. In this study, we have formulated an economical electroporation buffer for electroporation of RAW 264.7 cells compatible with commercial nucleofector apparatus. We observed that supplementation of membrane fusogenic agents such as Ficoll, PEG and membrane resealing agent, poloxamer P188, enhanced the transfection efficiency of macrophages to a level comparable to the commercially available solutions thereby providing us a cost effective solution for genetic manipulation of macrophages especially in large numbers. |
| Databáze: | OpenAIRE |
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