Sf9 Cells
General information
Description | Sf9 cells are clonal isolates derived from the Spodoptera frugiperda Sf21 cell line (IPLB-Sf-21-AE). They are commonly used in insect cell culture for recombinant protein production using baculovirus expression systems. Sf9 cells are epithelial in morphology and were cloned from the pupal ovarian tissue of the fall armyworm. One of the key characteristics of Sf9 cells is their small, regular size which is ideal for the formation of monolayers and plaques. They are also suitable for transfection, plaque assay/purification, amplification of high-titer stocks, and expression of recombinant proteins. The Sf9 insect cell line can be maintained in attached and suspended cultures, and do not require serum or CO2 to grow. They are considered Biosafety Level 1 and are usually grown in a 26-28 degree celsius incubator. Sf9 cells/baculovirus expression systems are widely used for high-level protein expression, often for purification, but proteins may also be functionally expressed in the defined Sf9 cell environment. The size of infected Sf9 cells is generally 17-30 microns in diameter. The Sf9 cell line is distinct from the Sf21 cell line in that it is a clonal isolate with a smaller and more regular size, while Sf21 cells are more disparate in size and form monolayers and plaques that are more irregular. Some Sf9 cell lines may harbor a negative sense Rhabdovirus called Spodoptera frugiperda rhabdovirus (SfRV), although not all tested Sf9 cells appear to be infected with this virus. The genome size of Sf9 has been estimated to be 451 Mbp with a G+C content of 36.53%. |
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Organism | Fall armyworm |
Tissue | Ovary |
Applications | Transfection, plaque assay/purification, amplification of high-titer stocks, and expression of recombinant proteins |
Synonyms | SF9, sf9, SF-9, Sf-9, sf-9, Sf 9, Spodoptera frugiperda clone 9, Sf clone 9, IPLB-Sf-9AE, IPLB-SF-9AE, IPLB-SF-9, IPLB-Sf-9, IPLB-Sf9 |
Characteristics
Age | Pupal stage |
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Gender | Female |
Morphology | Round, attached, epitheloid |
Growth properties | Monolayer, adherent |
Identifiers / Biosafety / Citation
Citation | Sf9 (Cytion catalog number 604328) |
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Biosafety level | 1 |
Expression / Mutation
Virus susceptibility | Baculoviruses, Autographa californica (MNPV), St. Louis encephalitis (SLE) |
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Handling
Culture Medium | Spodopan (PAN Biotech) |
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Medium supplements | Supplement the medium with 2% FBS to enhance proliferation if needed |
Passaging solution | Accutase |
Subculturing | Detachment of cells via a cell scraper is recommended. Collect the medium with detached cells after scraping in a 15ml centrifuge tube. Add about 5ml of medium to the flask and rinse the flask several times to collect any remaining cells and combine them with the rest of the cells in the tube. Centrifuge for 3 min at 300xg, remove the supernatant, resuspend the cells in fresh, cold medium and dispense into new flasks. |
Split ratio | For the first two subcultivations a ratio of 1:3 to 1:5 is recommended. In further subcultivations cells can be split at a ratio of 1:10 to 1:20 |
Seeding density | 1 x 10^4 cells/cm^2. Incubate between 26 to 30 degree Celsius in a non-humidified, ambient air-regulated incubator. Use cell culture flasks with filter caps or loosen caps to allow for oxygen exchange. |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
Amelogenin: x,x
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Required products
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.
Phosphate-buffered saline (PBS) is a versatile buffer solution used in many biological and chemical applications, as well as tissue processing. Our PBS solution is formulated with high-quality ingredients to ensure a constant pH during experiments. The osmolarity and ion concentrations of our PBS solution are matched to those of the human body, making it isotonic and non-toxic to most cells.
Composition of our PBS Solution
Our PBS solution is a pH-adjusted blend of ultrapure-grade phosphate buffers and saline solutions. At a 1X working concentration, it contains 137 mM NaCl, 2.7 mM KCl, 8 mM Na2HPO4, and 2 mM KH2PO4. We have chosen this composition based on CSHL protocols and Molecular cloning by Sambrook, which are well-established standards in the research community.
Applications of our PBS Solution
Our PBS solution is ideal for a wide range of applications in biological research. Its isotonic and non-toxic properties make it perfect for substance dilution and cell container rinsing. Our PBS solution with EDTA can also be used to disengage attached and clumped cells. However, it is important to note that divalent metals such as zinc cannot be added to PBS as this may result in precipitation. In such cases, Good's buffers are recommended. Moreover, our PBS solution has been shown to be an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, such as SARS-CoV-2.
Storage of our PBS Solution
Our PBS solution can be stored at room temperature, making it easy to use and access.
To sum up
In summary, our PBS solution is an essential component in many biological and chemical experiments. Its isotonic and non-toxic properties make it suitable for numerous applications, from cell culture to viral transport medium. By choosing our high-quality PBS solution, researchers can optimize their experiments and ensure accurate and reliable results.
Composition
Components
mg/L
Inorganic Salts
Potassium chloride
200,00
Potassium dihydrogen phosphate
200,00
Sodium chloride
8,000.00
di-Sodium hydrogen phosphate anhydrous
1,150.00