Table of Contents
How do I store my PEI solution?
Filter the PEI solution through a 0.22-μm nitrocellulose filter (e.g., GS filter, Millipore) and store the solution at 4°C. This 10% (w/v) solution is the working PEI reagent and appears to be stable indefinitely.
How do you store PEI Max?
Dissolve PEI in endotoxin-free dH2O that has been heated to ~80°C. Let cool to room temperature. Neutralize to pH 7.0, filter sterilize (0.22um), aliquot and store at -20°C; a working stock can be kept at 4°C.
Can PEI be frozen?
Furthermore, in contrast to other polyethylenimines, PEI F25-LMW-based DNA or siRNA complexes allow freeze/thawing and frozen storage for several months. Their activity is fully retained without requiring specific buffer conditions or the addition of any lyoprotectant.
Can PEI Max be frozen?
PEI-HCl-40K at 1mg/ml working solution: stable at 4 degree for up to 6 months. For long- term storage, put 3-5ml aliquots in -80 or -20 degree. Once thawed completely, filter through 0.22um filter, aliquot and keep at 4 degree. Do not freeze again.
How do you make Max PEI solution?
1. In 1L glass beaker, suspend 1g of PEI MAX 40K in 900 mL water. 2. Add PTFE-coated stir bar to 1L glass beaker and set stirring to produce small vortex.
How do you make a PEI transfection reagent?
Preparation
- Dissolve 100 mg in 100 mL sterile ddH2O.
- Stir while slowly adding HCl to pH 7.0.
- Mix for 10 minutes and then recheck pH.
- Filter sterilize through 0.22 um filter.
- Aliquot 500 uL to 1000 uL and store in -80C.
- Thawed solution can be stored at 4C for up to 2 months, label a tube when thawed.
What is PEI Max?
Polyethylenimine “Max” (PEI MAX) is a powerful, trusted, and cost-effective reagent widely considered as a current gold standard for both in vitro and in vivo transfection. PEI MAX has a high density of protonatable amino groups, with amino nitrogen as every third atom.
How do you thaw in PEI?
1: stir the PEI in MQ (1mg/ml) and dissolve by dropping and holding the pH at 2. After its transparant increase pH to 7 –> filter through 0.22um filter –> aliquot –> 4x freeze-thaw.
Why is PEI used for transfection?
DNA can be introduced into a host cell by transfection with polyethylenimine (PEI), a stable cationic polymer (Boussif et al., 1995). PEI condenses DNA into positively charged particles that bind to anionic cell surfaces. Our laboratory uses PEI over other cell transfection reagents because of its low cost.
How do you make PEI Max?
Why is PEI used in transfection?
How long is PEI transfection?
Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 15-20 min at room temperature.
Which is the best Pei transfection reagent for ATMP?
PEIpro ®-GMP is the highest quality grade PEI transfection reagent for use as raw material in the production of viral vectors (eg. AAV, lentivirus) intended for Human ATMP clinical trials and commercialization.
Which is the most effective transient transfection reagent?
PEI 25K is a powerful, trusted, and cost-effective transient transfection reagent. In HEK293 and CHO expression systems, PEI offers consistently high gene expression on a wide scale (96 well plates up to 100 L bioreactors). Each year, more researchers and companies turn to Polysciences PEI to gain a critical edge in their work.
Which is the best Pei for clinical use?
PEIpro ®-GMP is the highest quality grade PEI and the only transfection reagent available on the market supplied in a bag with MPC connectors and weldable tubing, ideal for GMP-compliant closed system manufacturing of clinical grade therapeutic viral vector, such as AAV and lentivirus.
Which is the best virus transfection reagent for peipro?
Irrespective of the cell culture-based system and production scale, PEIpro ® and PEIpro ® -HQ have led to efficient viral vector yields superior to 10 7 IG/mL and 10 9 VG/mL, respectively for lentiviruses and AAV (*to reach highest AAV titer yields in suspension cell culture systems, we recommend to use FectoVIR ® -AAV transfection reagent).