转染试剂 - 细胞生物学 - 中文站

产品中心

转染试剂

货号 品名 规格 价格
TR101 Clark转染试剂 1ml RMB 1290.00
订购

Clarkfection 是阳离子聚合物类型的转染试剂，相比脂质体类型具有更小的细胞毒性、更高的转染效率 以及更高的可重复性。
 

Clarkfection® is a superior cationic polymer-based transfection reagent. Compared with liposomes on the market, Clarkfection exhibits lower cytotoxicity, higher transfection efficiency and higher repeatability.

Clarkfection Features

  • For eukaryotic cells like HEK 293;
  • For both adherent and suspension cells;
  • For operations with or without serum;
  • More than 200,000 L application.

High protein expression and low cytotoxicity(EGFP protein expression transfected Clarkfection ）

 

293E 293FT
\ \ \ \
  COS7 HepG2
\ \ \ \
BHK21 MCF7
\ \ \ \
  SW480 CHO-DG44
\ \ \ \

Clarkfection Transfection Protocol

The transfection procedures of adherent cells are shown in Fig. 1.
we take the 96-well plates as example：
4cells/well (table 1)and cultured with 5%CO2 at 37℃ for 18-24h before transfection.
2. Preparation of transfection mixture:
(1). Dilute DNA into serum-free medium(25μL in total) and homogenize gently.
(2). Dilute Clarkfection into serum-free medium(25μL in total) and homogenize gently and incubate at room temperature for 5 min.
(3). Mix the DNA and Clarkfection at room temperature for 15~20min 3. Remove the culture medium and add 50μL mixture per well.
4. After 4-6hours(for SF9 cell is 2h), remove the transfection mixture and add medium with serum.
5. Gene expression is tested after incubation with 5% CO2 at 37℃ for 48-72h(incubate sf9 cell line at 27℃ for 48-72h).

 

 \

Fig. 1 Adherent cells' Transfection

Attention:

1). The experiment conditions for different cell lines are shown in Table 1. The corresponding transfection protocols can be got by entering links in Table 1.
(2). Number of cells per well, dosage of Clarkfection, DNA and serum-free medium for dilution are proportional to basal area per well for plates of different sizes. The basical areas for plates with different sizes are shown in Table 2. The dosage and ratio of DNA and Clarkfection should be optimized to achieve the best transfection results.

Table1. Usage of Clarkfection in different cell lines（96-well plate）

Cell type Culture medium Cells per well DNA Clarkfection Medium change after 4-6h
293H DMEM 3×104 0.2µg 0.5µL DMEM+10%FBS
293FT DMEM 3×104 0.2µg 0.5µL DMEM+10%FBS
293E DMEM 3×104 0.2µg 0.5µL DMEM+10%FBS
293F DMEM 3×104 0.2µg 0.5µL DMEM+10%FBS
COS7 DMEM 1.5×104 0.4µg 0.5µL DMEM+10%FBS
hela DMEM 2×104 0.3µg 0.5µL 1640+15%FBS
Caco2 MEM 3.5×104 0.3µg 0.75µL MEM+10%FBS
BHK21 MEM 2×104 0.2µg 0.5µL MEM+10%FBS
CHO-DG44 DMEM+HT+pro 2×104 0.5µg 0.5µL DMEM+HT+pro +10%FBS
RAW264.7 DMEM 3×104 0.2µg 0.5µL DMEM +10%FBS
MCF7 MEM/NEAA+0.01mg/mL insulin + sodium pyruvat 2×104 0.1µg 0.25µL MEM/NEAA+0.01mg/mL insulin + sodium pyruvat+10%FBS
SW480 IMDM 3×104 0.4µg 0.5µL IMDM +10%FBS
MDCK DMEM 4×104 0.6µg 1µL DMEM+10%FBS
CHO-K1 IMDM+Pro 3×104 0.2µg 0.5µL IMDM+Pro +10%FBS
HepG2 DMEM 3×104 0.5µg 0.75µL DMEM+10%FBS
A549 DMEM 2×104 0.3µg 0.5µL DMEM+10%FBS
NIH/3T3 DMEM 1.5×104 0.1µg 0.75µL DMEM+10%FBS
vero DMEM 3×104 0.3µg 0.75µL DMEM+10%FBS
sf9 SIM SF 5×104 0.4µg 0.75µL SIM SF+10%FBS

Table 2. Scaling up or down transfections with Clarkfection（according to the growth area）.
Note: the actual conditions should be optimized by experiments.

\

top