Product Details:
Payment & Shipping Terms:
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Method-1: | Transient Transfection | Method-2: | Stable Transfection |
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Customer Offer: | Gene Sequences | Starting From: | Gene Synthesis |
Protein Purification: | Available | Report: | QC Report |
High Light: | QC Report Mammalian Expression System,Assemble Proteins Mammalian Expression System |
Introduction
Mammalian cells can correctly and effectively express eukaryotic proteins, including the synthesis, processing, and secretion of proteins. The mammalian system has the excellent function of post-translational modification of proteins and can assemble proteins correctly. This is because mammalian systems have endoplasmic reticulum, Golgi apparatus and other organelles that are key elements in cell transport and post-translational modification.
Most post translational modifications occur during or shortly after translation. Excreted proteins are made in the endoplasmic reticulum where there are enzymes that facilitate proper folding, disulfide bonding, and N-linked glycosylation. Additionally, before it is excreted, a protein would pass through the golgi apparatus, where O-linked glycosylation may occur as well. All of these modifications occur in many proteins expressed by mammalian cells, including antibodies.
There are two ways for protein expression of mammalian cells: transient transfection and stable transfection. HEK293 or CHO cells are often used in transient transfection. A small amount of protein can be quickly obtained within one week after the transient transfection. Meanwhile, there are dozens of cells that can be stably transfected, which can be used to produce cell lines after stable transfection and screening. Stable cell lines can grow continuously over a long period of time without significant change in the expression level of the target gene.
Experimental Process
Component |
Transient Transfection |
Stable Transfection (For Research Assay) |
Stable Transfection |
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Plasmid | supercoiled DNA | linear DNA | linear DNA |
Transfection cell | HEK293, CHO | CHO, HEK293, HeLa, M14, THP-1, BHK21, HFF-1, HepG2, MCF, Vero, AGS and more | CHO |
Turnaround | ~4 weeks | ~2 months | ~5 months |
Cost | + | ++ | +++ |
Application |
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Transfection method | Microinjection, electroporation, liposome transfection, calcium phosphate method, retroviral transfection |
Transient Transfection
Quickly transform cloned genes into large amounts of protein in 3-7 days.
Obtain fully post-translational modified and active mammalian protein.
Easily purify secreted protein from serum-free cell culture medium.
Expand culture volume from that of 50 multi-well plates to that of 25L and more
Stable Transfection