Chariot™* is a revolutionary delivery reagent that quickly and efficiently transports biologically active proteins, peptides and antibodies directly into cells. The typical delivery efficiency is 60-95%. Less than two hours after delivery, live cells can be assayed to determine the effects of the introduced material, without the need for fixing. This makes Chariot an ideal tool for functional studies, including delivering inhibitory proteins, labeling organelles, screening peptide libraries and studying protein half-lives & transient complementation.
The beta-Galactosidase Staining Kit has been DISCONTINUED. However, customers can use any commercially available beta-Galactosidase Staining Kit to determine the transfection efficiency of the positive control protein. For example: Invitrogen beta-Gal Staining Kit, catalog number: K146501.
|β-Galactosidase Staining Kit||75 rxns||35001||Discontinued|
|β-Galactosidase Staining Kit Manual|
|Cell Biology Products Brochure|
- Works in a variety of cell lines (see table on the User Data tab)
- Facilitates functional studies in living cells — no fixing required
- Works in vivo1
- Fast and efficient — 60-95% transfection in less than 2 hours
- No need for expression of fusion proteins
- Non-cytotoxic, serum independent
Why use Chariot™?
Current transfection techniques include cationic liposomes2, calcium phosphate coprecipitation3, electroporation4, microinjection5 and viral vectors6. These methods are useful but are very time-consuming as once transfection is completed, it takes 12-80 hours to detect expression of the gene of interest. A greater drawback is that the cytotoxicity of these methods can make it difficult to determine if any observed changes are caused by the recombinant protein or induced by the transfection method itself.
Chariot is a novel transfection method that eliminates many of the problems encountered with more commonly used procedures. The Chariot peptide forms a non-covalent bond with the macromolecule of interest. This stabilizes the protein, protecting it from degradation and preserving its natural characteristics during the transfection process7, 8. After delivery, the complex dissociates, leaving the macromolecule biologically active and free to proceed to its target organelle. Delivery occurs in the presence or absence of serum and is independent of the endosomal pathway, which can modify macromolecules during internalization. This, coupled with the fact that Chariot is non-cytotoxic, ensures that observed changes are the result of your recombinant protein, and not the transfection method.
Diagram 1: Flow chart of the Chariot process.
Chariot has been used to efficiently transfect a variety of cell lines. As a general rule, the larger the protein to be delivered, the slower it will be delivered; increasing protein size may also reduce delivery efficiency, though increasing delivery time will usually allow for adequate delivery. In contrast, Chariot cannot deliver very small peptides because they do not form a viable complex with the Chariot peptide. Therefore, we recommend that peptides be a minimum of 10 amino acids in length.
Chariot™ delivers for you
The ability of Chariot to deliver antibodies was demonstrated using FITC-labeled Actin and Lamp-1 antibodies. These were targeted to Actin filaments (Figure 1) and endosomes (Figure 2), respectively.
Figure 1: Delivery of fluorescent Actin antibody.
Figure 2: Delivery of fluorescent Lamp-1 antibody.
β-galactosidase is a protein that is composed of four identical subunits. The ability of Chariot to deliver protein was demonstrated using a 119 kDa subunit of β-galactosidase (Figure 3).
Figure 3: Delivery of β-galactosidase into HeLa cells.
The non-covalent Chariot-macromolecule complex dissolves after delivery, leaving the macromolecule biologically active and free to proceed to its target organelle, if it contains specific signaling sequences. This was demonstrated using a 51-mer nuclear protein (Figure 4) and a 32-mer cytoplasmic peptide (Figure 5).
Figure 4: Nuclear-targeted protein delivery.
Figure 5: Cytoplasmic peptide delivery.
Chariot speeds and simplifies a variety of functional studies because it efficiently delivers biologically active proteins, peptides and antibodies directly into live mammalian cells. This means you’ll get conclusive data immediately, rather than having to wait days for ambiguous expression data. For additional information, please download the Chariot manual and product profile.
- Aoshiba, K. et al. (2003) Am. J. Resp. Cell & Mol. Biol. 28: 555-562.
- Felgner, P.L. et al. (1987) Proc. Natl. Acad. Sci. USA 84: 7413-7417.
- Graham, F.L. and van der Eb, A.J. (1973) Virology 52: 456-467.
- Neumann, E. et al. (1982) EMBO J. 7: 841-845.
- Capecchi, M.R. (1980) Cell 22: 479-488.
- Cepko, C.L. et al. (1984) Cell 37: 1053-1062.
- Morris, M. et al. (1999) J. Biol. Chem. 274 (35): 24941-24946.
- Morris, M. et al. (2001) Nature Biotech 19: 1173-1176.
Chariot’s ability to effectively deliver proteins, peptides and antibodies into a variety of cell types is well documented. To download a list of nearly 100 publications citing the use of Chariot, click here.
The following table is compiled from data furnished to us by our customers. This chart is by no means exhaustive.
|Cell line/model||Cell types||Macromolecule delivered||Transfection efficiency|
|293||Human embryonic kidney||GFP||80%|
|A549||Human lung carcinoma||Antibody||95%|
|Arabidopsis||Primary plant protoplasts||Protein||Not provided|
|COS-7||Monkey kidney||Protein, oligopeptide||80%|
|Embryonic cells||Primary mouse||Protein||90%|
|Fetal Endothelial cells||Primary mouse||Protein||70-80%|
|HeLa||Human cervix carcinoma||Protein, peptide, antibody||95%|
|HMSC||Primary human mesenchymal stem cells||Peptide||80%|
|HS-68||Human foreskin fibroblast||Protein, peptide, antibody||95%|
|IMR90||Human fibroblast||Protein, peptide||80%|
|Jurkat||Human T-cell leukemia||Protein, peptide, antibody||75%|
|MCF-7||Human breast carcinoma||GFP||70%|
|Mono Mac 6||Primary human monocyte||Protein||50%|
|Mouse hepatocytes||Primary liver||Protein||95%|
|Mouse model (in vivo)||Alveolar wall tissue||Protein||88%|
|Neural retina cells||Primary chicken||Protein||80%|
|NIH/3T3||Mouse embryo||Protein, peptide||80%|
|NRK||Normal rat kidney||Antibody||30-60%|
|Pancreatic cells||Primary human||Protein||90%|
|PC-12||Rat pheochromocytoma||Protein, peptide||80%|
|Saos-2||Human osteosarcoma cells||Antibody||50-70%|
|SCC-61||Primary human head + neck tumor||Peptide||90%|
|Sensory neurons (DRG)||Primary chicken||Antibody, peptide||80%|
|Sensory neurons (DRG)||Primary rat||Protein||80%|
|WI-38||Human lung fibroblast||Protein, peptide, antibody||95%|
|WISH||Human placenta carcinoma||Protein||30%|
Contents & Storage
Chariot is offered in 25- and 100-reaction kits, where a reaction is defined as the amount of reagents required to deliver the β-galactosidase positive control protein to cells in one well of a 6-well plate or in a 35 mm plate. The number of reactions you can perform will vary depending on the size plates used and the type of macromolecule(s) delivered. Each Chariot Kit contains sufficient Chariot Transfection Reagent, PBS, β-galactosidase positive control protein and sterile water for either 25 or 100 reactions. Store all reagents at -20°C. All reagents are guaranteed stable for 6 months.
Some select publications are listed below.
- “Inhibition of Tunneling Nanotube (TNT) Formation and Human T-cell Leukemia Virus Type 1 (HTLV-1) Transmission by Cytarabine.” by Omsland et al. (2018) Scientific Reports 8(11118).
- “Co-dependence of HTLV-1 p12 and p8 Functions in Virus Persistence.” by Pise-Masison et al. (2014) PLoS Pathogens 10(11): e1004454.
- “Mouse embryonic stem cells inhibit murine cytomegalovirus infection through a multi-step process.” by Kawasaki et al. (2011) PLoS One 6(3): e17492.