Transfection Reagent
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一、Product Introduction:
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Product Name: Transfection Reagent;
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Abbreviation: TR
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Cat. No:FH880806
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Shelf life : 18 months;
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Specification: Concentration 1 µg/ µL; Volume 100ul / 750ul / 1.5ml；
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Storage conditions:
It is recommended to store at 2-8℃ after aliquoting, and do not store at -20℃
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二、Product Description：
TR is a polycationic polymer with high cationic charge density potential that can act as an effective "proton sponge" at any pH. This polymer can transfer different reporter genes into various cells, and its effect is better than lipopolyamide. After further modification, its transfection performance is better than dendrimers, and its cytotoxicity is lower.
三、Protocol：
The ratio of DNA (µg) to TR (µg) used for most cell lines is 1:3 or 1:4. Cells with good transfection status can obtain high transfection efficiency, high expression level and low cytotoxicity.
Take the 24-well plate as an example:
3.1 Adherent cells: One day prior to the transfection, 0.5-2×10E5 cells are seeded per well in . The cells can grow to 90-95% confluency at the time of transfection.
Suspension cells: 4-8×10E5 cells are seeded per well in 500 µL of complete medium prior to preparation of TR transfection solutions.
3.2 Preparation of TR transfection solutions as follows:
A. Dilute the plasmid DNA in 50 µL of DMEM(or other serum-free medium). Mix gently.
B. Dilute the appropriate amount of TR in 50µL of DMEM. Incubate for 5 minutes at room temperature.
C. Combine the diluted DNA with diluted TR. Mix gently and incubate for 30 minutes at room temperature.
3.3 Add the 100ul of DNA-TR complexes to a well containing cells and
medium. Mix gently.
3.4 For adherent cells, the medium can be changed 4-6h after transfection, while for suspension cells, the medium can be changed 18-48h after transfection. Gene expression can be detected 18-48h after transfection. If detecting protein expression, please collect cell samples 24-72h after transfection.
3.5 For stable transfection, passage cells at a 1:10 into complete medium 24 hours after transfection. Add selective medium the following day.
四、Examples-The transfection efficiency of TR and other transfection reagent in HEK293 & HepG2：
