HiPerFect Transfection Reagent

The composition is confidential.   HTS is a different component from HiPerFect.  HTS works with very low amounts of reagent for transfection, and is specifically adapted for use in high throughput applications.  HiPerFect Transfection Reagent is for siRNA transfection at lower throughput applications.

For the fixation of cells transfected with fluorescently labeled siRNA, we would suggest to perform the following protocol:

1. After transfection, remove the medium from the cells and wash the cells once with PBS.
2. Incubate the cells for 15 minutes at room temperature with 4% Paraformaldehyde (in PBS, pH 7.0). The cells should be completely covered by this solution (e.g., for a 96-well plate use 50 µl solution/well)
3. Wash the cells with PBS.

Fixed cells can be stored at 4°C for a few days.

(Note: It is also possible to use chamber slides or object slides for this procedure. Object slides should be coated to provide better growing conditions for cells. Cells can be fixed as described above and then overlayed with embedding medium to allow investigation using a fluorescence microscope. Optimal conditions for this method need to be determined by the user)

 

Yes, please follow the protocol "Transfection of Suspension Cell Lines, including Jurkat and K562, with siRNA in 24-Well Plates" in the HiPerFect Transfection Reagent Handbook.

 

 

Unfortunately, we do not have any data for the transfection of siRNA into insect cells. However, we know about customers who have successfully used Effectene Transfection Reagent for the transfection of plasmid DNA into insect cell lines S2 and Sf9 (see FAQ 397). Since Effectene Transfection Reagent and HiPerFect Transfection Reagent for the transfection of eukaryotic cells with siRNA are both lipid based, there is a chance that HiPerFect will work with insect cells. It is especially suitable for transfection of very low siRNA concentrations, reducing the risk of cell toxicity and off-target knockdown effects.

Alternatively, you can try RNAifect or TransMessenger Transfection Reagent, also lipid-based, for the transfection of siRNA into S2 cells.

We would appreciate any feedback on your results in case you want to give it a try. Please visit our Transfection Cell Survey page to submit your feedback.

Yes, please follow the protocol "Transfection of Macrophage Cell Lines, Including J774.A1 and RAW 264.7, with siRNA in 24-Well Plates" in the HiPerFect Transfection Reagent Handbook.

 

 

The amount of HiPerfect Transfection Reagent and siRNA required for optimal performance may vary, depending on the cell line and gene target. When transfecting fluorescently labeled siRNA (e.g., Alexa Fluor 488 siRNA) for monitoring transfection efficiency under a fluorescent microscope, it may be necessary to increase the amount of siRNA up to 25 nM (without increasing the volume of HiPerFect Reagent), depending on the optical requirements of the microscopic equipment that will be used for detection.

Yes, please follow the protocol 'Reverse Transfection of Adherent Cells with siRNA in 384-Well Plates' in the HiPerFect Transfection Reagent Handbook.

 

HiPerFect Transfection Reagent does work for the transfection of miScript miRNA Mimics and Inhibitors, and the new handbook contains protocols for this application.

 

 

HiPerFect Transfection Reagent is optimized for siRNA transfections and enables effective siRNA uptake and efficient release of siRNA inside cells, resulting in high gene knockdown even when using low siRNA concentrations. For high-throughput siRNA screenings HiPerFect HTS Reagent is a fast and effective newcomer specifically designed to focus on robustness and cost efficiency. For the transfection of shRNA.

Attractene Transfection Reagent should be used for the transfection of shRNA (short-hairpin RNA) vectors for gene silencing experiments to achieve high efficiency. Ease and flexibility of handling enables preparation and storage of transfection complexes making Attractene Reagent suitable for use with automated systems.

http://www.sabiosciences.com/reversetransfection.php

The Molecular Weight (MW) of a 21 nucleotide double-stranded siRNA molecule is approximately 13-15 ug/nmol. The exact MW depends on the sequence of the siRNA. 20 uM of double-stranded 21 nt siRNA is equivalent to approximately 0.25 ug/ul.