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Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for C6 Cells (Glioma Cells, CCL-107) Altogen Biosystems offers the C6 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The C6 Transfection Reagent is an advanced lipid formulation reagent, and it has been developed to provide high transfection efficiency with the C6 cell line. This cell line is a good host for studying mammalian brain tumors and cell and molecular biology. When cultured in vitro, C6 cells are a monolayer and adhere to the surface of the culture flask. Other applications include drug discovery, gene expression studies, molecular and cell biology research applications. Purchase C6 Transfection Kit at www.Altogen.com Altogen Biosystems 848 Rainbow Blvd #823 Las Vegas NV 89107 USA Telephone 702 349 6103 Fax 702 989-0841 email [email protected] Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for C6 Cells (Glioma Cells, CCL-107) C6 Cell Line Description and Applications C6 cells The C6 cell line originates from the glial tumor from the brain tissue of a rat (Rattus norvegicus) and induced by Nnitrosomethylurea. It was cloned and isolated by Benda et al. after several passages. The C6 cell line displays fibroblast morphology as well as glucocorticoid receptors. The C6 cell line can provide a good transfection model for research, but it can be particularly useful for research related to mammalian brain tumors. The C6 stem line number is diploid and the karyotype is stable. Altogen Biosystems 848 Rainbow Blvd #823 Las Vegas NV 89107 USA Telephone 702 349 6103 Fax 702 989-0841 email [email protected] Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for C6 Cells (Glioma Cells, CCL-107) C6 Transfection Protocol 1. Plate 10,000 - 15,000 C6 cells per well in 0.5 ml of complete growth medium 12–24 hours prior to transfection 2. Wash with 1xPBS and add 0.5 ml of fresh growth medium 3. Prepare transfection complexes by mixing 40 µl of serum-free medium, 5.5 µl of transfection reagent, and • 750 ng DNA (or mRNA), or • 30 nM - 50 nM of siRNA (or microRNA) *Referred to a final volume including growth medium 4. Incubate transfection complexes at RT for 15 - 30 minutes 5. Optional: Add 2 µl of Complex Condenser. This reagent reduces the size of transfection complex, therefore increasing transfection efficiency; however it may increase cell toxicity 6. Add prepared transfection complexes to 0.5 ml of complete growth medium with C6 cells (from step 2) 7. Incubate cells at 37ºC in a humidified CO2 incubator 8. Assay for phenotype or target gene expression 48 - 72 hours after transfection Optional: Transfection efficiency can be increased by addition of Transfection Enhancer reagent. Add 2 µl of Transfection Enhancer reagent 12-24 hours after transfection If the viability of C6 cells being transfected is affected at 16 - 24 hours post-transfection, the level of cytotoxicity can be decreased by changing the growth medium and eliminating redundant exposure of cells to transfectant Altogen Biosystems 848 Rainbow Blvd #823 Las Vegas NV 89107 USA Telephone 702 349 6103 Fax 702 989-0841 email [email protected] Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for C6 Cells (Glioma Cells, CCL-107) General Lipoplex-mediated Transfection Mechanism of Action Altogen Biosystems 848 Rainbow Blvd #823 Las Vegas NV 89107 USA Telephone 702 349 6103 Fax 702 989-0841 email [email protected] Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for C6 Cells (Glioma Cells, CCL-107) C6 Transfection Kit Product Details •Two component formulation enhances lipid mediated transfection efficiency •Transfection protocols provided for transfection of proteins, DNA, mRNA, siRNA, shRNA and microRNA •Transfection Enhancer reagent provided with the kit •Produce higher level of recombinant protein expression with minimal disruption of normal cell function •Generate physiologically relevant data you can trust •Effective for plasmid DNA/siRNA co-transfection •Easy-to-use transfection protocol with reproducible results •Low cytotoxicity Developed and manufactured by Altogen Biosystems (www.Altogen.com) Altogen Biosystems 848 Rainbow Blvd #823 Las Vegas NV 89107 USA Telephone 702 349 6103 Fax 702 989-0841 email [email protected] Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for C6 Cells (Glioma Cells, CCL-107) Data Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into C6 cells following the recommended protocol. At 48 hours post-transfection the cells were analyzed by qRT-PCR for Lamin A/C gene expression levels. 18S rRNA levels were used to normalize the Lamin A/C data. Values are normalized to untreated sample. Data are means ± SD (n=4). Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for C6 Cells (Glioma Cells, CCL-107) Data Figure 2. Protein expression of Lamin A/C in C6 cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into C6 cells following Altogen Biosystems transfection protocol. At 72 hours posttransfection the cells were analyzed by Western Blot for protein expression levels (normalized by total protein, 10 µg of total protein loaded per each well). Untreated cells used as a negative control. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for C6 Cells (Glioma Cells, CCL-107) C6 Transfection Kit Benefits • Pre-optimized transfection protocol for C6 cell line • Compatible with DNA, small RNA (siRNA, shRNA, miRNA), mRNA, and small protein complexing • Free of serum and protein of animal origin • Compatible with standard and reverse transfection methods (both protocols provided in the kit manual) • Easy to use C6 transfection protocol ensures great performance with expedited experimental timeline • Equally efficient for single or multiple transfections • Can be used for transient transfection and development of stable C6 cell lines • Bio-degradable after endocytosis • Used for preclinical research worldwide Altogen Biosystems 848 Rainbow Blvd #823 Las Vegas NV 89107 USA Telephone 702 349 6103 Fax 702 989-0841 email [email protected]