Contents of the Kit:
PT-R
| :1 tube |
| Lipoform buffer | :0.2 ml |
| Enhancer E | :1 tube
|
Storage Condition : 0-5ºC
Shipping Condition: ambient temperature
Required Equipment and Materials
10 μl, 100-200 μl, and 1000 μl pipettes; CO2 incubator, microtubes (sterile)
Product Description
CarryMax-R is a protein delivery kit designed to achieve the best performance of protein delivery to various animal cells. CarryMax-R’s protein delivery mechanism is similar to gene delivery by cationic liposomes. The protein-cationic liposome complex binds on the cell membrane and then the complex is taken inside cells by endocytosis; upon entry, the protein is released to the cytoplasm. The location of the protein delivered by this method will be determined by various experiments.
Recent studies have shown that the delivered protein in the cell, such as apoptosis related proteins, works in the same manner as a protein synthesized in the cell by gene expression. This delivery method will be very useful in understanding the function of the protein in the cell. The total time to deliver proteins is about 2-4 hours if no further cell culture is required. Therefore, less time-consuming experiments will be possible to check the function of the protein than when using a DNA transfection method. Protein delivery techniques are not an alternative method to DNA transfection Techniques; rather, they are complementary to each other. Both techniques can be used together, depending on the purpose of the experiment.
CarryMax-R can be applied to a wide variety of studies related to intracellular protein functions, such as apoptosis, cell cycle regulation, cell signaling, DNA transcription, protein expression regulation, and RNA splicing. Additionally, since antibodies or other foreign molecules, such as peptides, biomolecules, and charged polymers, can be delivered into a viable cell, therapeutic applications of CarryMax will be possible. In the near future, dual delivery of protein and DNA by CarryMax-R will be another possible application.
General Protocol
Preparation of protein delivery solution
1. Add 0.2 ml of Lipoform Buffer to the PT-R tube and dissolve with a vortex mixer for 30 seconds.
2. Add 40 μl DMSO to Enhancer E and dissolve by pipetting.
3. Mix 100 μl of PT-R solution and 5 μl of Enhancer E solution in a microtube followed by vortexing for 10 seconds to prepare Liposome solution.
Preparation of Protein-Protein delivery reagent complex1. Add 30 μl of sterilized water solution to the tube.
2. Add 1 μg protein and mix with pipetting.
3. Add 4 μl Protein delivery solution to the tube and mix with pipetting.
4. Incubate for 15 minutes at room temperature.
Transfect to cells
1. Change the cell culture media to fresh, serum-free media.
2. Add Protein delivery reagent complex solution to the cells.
3. Change the culture media to media containing serum 4 hours after the addition of the Protein delivery complex solution.
4. Continue to incubate for another 4 hours or appropriate period of time depending on the experiment.