Log In Sign up
The largest database of trusted experimental protocols

Transit 2020 kit

Manufactured by Mirus Bio
Visit Supplier

The TransIT-2020 kit is a reagent-based system designed for the efficient transfection of nucleic acids, such as plasmids, into a variety of cell types. The kit provides the necessary components to facilitate the delivery of genetic material into cells, enabling researchers to study gene expression, protein production, and other cellular processes.

Automatically generated - may contain errors

Lab products found in correlation

In fusion kit, by Takara Bio (2 mentions) P adeno x zsgreen1 vector, by Takara Bio (1 mentions)

Spelling variants of this product

TransIT-2020 (151 citations)

3 protocols using transit 2020 kit

1

Overexpression of ESRP2 in HepG2 and AML12 cells

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
HepG2 and AML12 cell lines were obtained from ATCC (Catalogue no. HB-8065 and CRL-2254, respectively) and cultured according to specifications as stated by ATCC. When tested for mycoplasma contamination (Biotool, catalogue no. B39032) prior to use, these cell lines tested negative. cDNAs encoding FLAG-tagged mouse Esrp2 (ref. 43 (link)) were sub-cloned into the p-Adeno-X-ZsGreen1 vector (Clonetech, 632267) using the In-Fusion kit (Clonetech, 639646) as per the manufacturer's instructions. High-titre adenoviruses were generated by transfecting Ad-293 cells (∼70% confluent) in T-25 flasks with linearized recombinant adenoviral plasmid using Mirus TransIT-2020 kit and cells were harvested once cytopathic effect was seen. Following this, two viral amplification steps were performed and the viral particles were purified using CsCl gradient as mentioned in the Adeno-X Adenoviral System 3 user manual. After purification of viral particles, the titre was determined by ultraviolet spectrophotometry at 260 nm. To determine the effect of overexpression of ESRP2 in HepG2 and AML12 cells, T-25 flasks containing ∼50% confluent HepG2 and AML12 cells were infected with 1.5 × 109 o.p.u. (optical particle units) of the ESRP2 or GFP adenovirus for 48 h and cells were harvested to extract RNA and protein for further analysis.
Bhate A., Parker D.J., Bebee T.W., Ahn J., Arif W., Rashan E.H., Chorghade S., Chau A., Lee J.H., Anakk S., Carstens R.P., Xiao X, & Kalsotra A. (2015). ESRP2 controls an adult splicing programme in hepatocytes to support postnatal liver maturation. Nature Communications, 6, 8768.
+ Open protocol
+ Expand
2

Overexpression of ESRP2 in HepG2 and AML12 cells

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
HepG2 and AML12 cell lines were obtained from ATCC (Catalogue no. HB-8065 and CRL-2254, respectively) and cultured according to specifications as stated by ATCC. When tested for mycoplasma contamination (Biotool, catalogue no. B39032) prior to use, these cell lines tested negative. cDNAs encoding FLAG-tagged mouse Esrp243 (link) were sub-cloned into the p-AdenoX-ZsGreen1 vector (Clonetech, 632267) using the InFusion kit (Clonetech, 639646) as per the manufacturer’s instructions. High-titer adenoviruses were generated by transfecting Ad-293 cells (~ 70% confluent) in T-25 flasks with linearized recombinant adenoviral plasmid using Mirus TransIT-2020 kit and cells were harvested once cytopathic effect (CPE) was seen. Following this, two viral amplification steps were performed and the viral particles were purified using CsCl gradient as mentioned in the Adeno-X™ Adenoviral System 3 user manual. After purification of viral particles, the titer was determined by UV spectrophotometry at 260 nm. To determine the effect of overexpression of ESRP2 in HepG2 and AML12 cells, T-25 flasks containing ~50% confluent HepG2 and AML12 cells were infected with 1.5 X 109 opu (optical particle units) of the ESRP2 or GFP adenovirus for 48 hours and cells were harvested to extract RNA and protein for further analysis.
Bhate A., Parker D.J., Bebee T.W., Ahn J., Arif W., Rashan E.H., Chorghade S., Chau A., Lee J.H., Anakk S., Carstens R.P., Xiao X, & Kalsotra A. (2015). ESRP2 controls an adult splicing programme in hepatocytes to support postnatal liver maturation. Nature Communications, 6, 8768.
+ Open protocol
+ Expand
3

Transient Transfection and MMP9 Overexpression

Check if the same lab product or an alternative is used in the 5 most similar protocols
For transient transfection experiment, we used pCMV empty vector and pCVM vector containing full‐length AR. For induction of MMP9 overexpression into the PCa cells, PLX‐304 empty vector and PLX‐304‐MMP9 vector were used. TransIT‐2020 kit (Mirus Bio, MIR5410, Madison, WI) was used by following the manufacturer's protocol.
Larsson P., Syed Khaja A.S., Semenas J., Wang T., Sarwar M., Dizeyi N., Simoulis A., Hedblom A., Wai S.N., Ødum N, & Persson J.L. (2019). The functional interlink between AR and MMP9/VEGF signaling axis is mediated through PIP5K1α/pAKT in prostate cancer. International Journal of Cancer, 146(6), 1686-1699.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!