Why transfect purified proteins directly?

Protein transfection is extremely rapid compared to traditional gene expression studies using transfected DNA (1–2 hours compared to 18–48 hours) because it bypasses the cellular processes of transcription and translation. It also facilitates studies involving transient effects of proteins, and avoids potentially harmful, random DNA integration into the genome of the target cells. Xfect Protein [...]

What is Xfect Protein Transfection Reagent and how does it work?

Xfect Protein Transfection Reagent is a modified peptide with a cell-penetrating activity whose amino acid composition enables it to interact with a protein cargo and transport this protein across the cell membrane barrier. Just incubate your protein of interest with Xfect in the supplied buffer for 30 min, then apply the mixture to your cells—2 hours later, you are ready to assay for protein [...]

How to assess transfection efficiency after transfection?

Reporter assays: you can use a variety of methods to measure transfection efficiency, including fluorescent and non-fluorescent reporter assays, qRT-PCR, and Western blotting. Please note that some assays are more rapid and sensitive than others. When to run your assay: most reporter assays are performed 1–3 days after transfection. The exact timing will depend on the type of cells you are [...]

How long should transfection duration be?

The optimal incubation period for cargo and reagent varies with each transfection experiment and must be determined empirically. Since prolonged exposure to transfection reagents may result in cell toxicity, it is critical to stay within the time range recommendations advised in your manual.Transfection [...]

What is the optimal plasmid quality for transfection?

Use only high-purity DNA (OD260/280 >1.8) for plasmid transfection. We recommend Macherey-Nagel NucleoBond Xtra Midiprep and Maxiprep kits—including the endotoxin-free kits—for transfection-grade plasmid purification.Transfection [...]