FAQ - NeuroPORTER Transfection Reagent

FAQs: NeuroPORTER™ Transfection Reagent

1. What is the NeuroPORTER Transfection Reagent?
2. Is the NeuroPORTER Transfection Reagent recommended for transfection of siRNA into primary neurons?
3. What media was used for the transfection of rat primary cortical neurons shown on the NeuroPORTER webpage?
4. What procedure is recommended for immunofluorescent antibody detection of transgenic proteins expressed in primary neurons?
5. How stable is the NeuroPORTER Transfection Reagent?

1. What is the NeuroPORTER Transfection Reagent?

The NeuroPORTER Transfection Reagent is a unique cationic lipid specifically formulated for efficient DNA transfection into neuronal cell lines, glial cells, and certain primary neurons.

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2. Is the NeuroPORTER Transfection Reagent recommended for transfection of siRNA into primary neurons?

For transfection of siRNA into primary neurons, we recommend using the GeneSilencer® siRNA Transfection Reagent. Click here to see more information about GeneSilencer® siRNA Transfection Reagent.

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3. What media was used for the transfection of rat primary cortical neurons shown on the NeuroPORTER webpage?

The following medium was used:

• 500 ML DMEM/ HIGH GLUCOSE (sigma D5671)
• 62.5 ML BOVINE CALF SERUM(defined hight and inactivate) (Hyclone)
• 1,5 ML PEN/STREP (Sigma P-0781)
• 6.25 ML GLUTAMINE(200 mM) (Sigma G7513)
• 15.6 ML HEPES (Omega HB-20)
• 62.5 ML MEDIA F-12 (Sigma F4888)

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4. What procedure is recommended for immunofluorescent antibody detection of transgenic proteins expressed in primary neurons?

The following procedure was used for immunofluorescent antibody detection in primary rat cortical neurons:

• Wash cover slips 3 times with PBS-BSA.
• Permeabilize cells with PBS-BSA + TritonX 0.1% for 5-10 minutes at room temperature (RT).
• Wash 1 time with PBS-BSA.
• Add Primary Antibody diluted in PBS-BSA, TrintonX 0.05% to cells.
• Incubate for 1 hour at RT.
• Wash 3 times with PBS-BSA + TritonX 0.05%.
• Add Secondary Antibody diluted in PBS-BSA, TrintonX 0.05% to cells.
• Incubate for 1 hour at RT.
• Wash 2 times with PBS-BSA
• Wash 3 times with PBS.
• Mount cover slips using media specific for fluorescence.

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5. How stable is the NeuroPORTER Transfection Reagent?

All components are stable for at least one year if stored properly.