Attractene Transfection Reagent

shRNA gene silencing Human - HEK 293T CAPN5- (Calpains) cationic lipid based

Experiment
shRNA gene silencing Human - HEK 293T CAPN5- (Calpains) cationic lipid based
Product
Attractene Transfection Reagent from Qiagen
Manufacturer
Qiagen

Protocol tips

Upstream tips
Seed 5.0 × 10^5 cells
Protocol tips
Add 4.5 μL Attractene Transfection Reagent to the DNA solution.

Incubate the cells under their normal growth conditions and change medium after 6h and continue to incubate until 48 h

Publication protocol

HEK293T-CAPN5-p.R243L cells were transfected using Attractene Transfection Reagent (Qiagen). In each well of 6-well plates, 5.0 × 105 HEK293T-CAPN5-p.R243L cells were grown in 2 mL complete DMEM medium to 50 - 70% confluence. Growth medium was replaced with 2 mL of freshly prepared Opti-MEM medium containing 5% FBS, penicillin (100U/mL) and streptomycin (100 μg/mL) (complete Opti-MEM medium). Cells were then transfected with 3.0 μg shRNA plasmid using 4.5 μL Attractene Reagent. At 6 hours post-transfection, growth medium was replaced with fresh complete Opti-MEM medium. Cells were incubated for 48 hours.

The optimization protocols (varying pulse voltage, pulse width and pulse, as recommended by manufacturer) for transfecting adherent and suspension cells with Neon™electroporation system (Invitrogen; Carlsbad, CA) were performed with SH-SY5Y cells. Briefly, 1.0 × 106 SH-SY5Y cells in 100 μL resuspension buffer were electroporated with 12.0 μg shRNA plasmids into 150 × 20 mm tissue culture dishes containing 15 mL Opti-MEM medium containing 5% FBS. After optimization, transfection of SY-SY5Y cells were performed in triplicate for each shRNA plasmid and incubated for 72 hours.

Full paper   Login or join for free to view the full paper.

Reviews

Attractene Transfection Reagent from Qiagen has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Discussion

5 years ago

Author: Italy

Is a knockdown using shRNA permanent?

Is a knockdown using shRNA permanent and if not is there a known duration?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing shRNA gene silencing Human - HEK 293T CAPN5- (Calpains) cationic lipid based using Attractene Transfection Reagent from Qiagen.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Qiagen for Attractene Transfection Reagent below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing shRNA gene silencing Human - HEK 293T CAPN5- (Calpains) cationic lipid based using Attractene Transfection Reagent from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms