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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 COLO-205 Cells (Colon Adenocarcinoma Cell, CCL-222) Altogen Biosystems offers the COLO-205 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The COLO-205 Transfection Reagent is an advanced nanoparticle-based liposome formulated transfection reagent that enhances lipid mediated transfection, and it has been developed to provide high transfection efficiency with COLO-205 cells. This cell line is a good host for studying colon cancer. When cultured in vitro, COLO-205 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 COLO-205 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 COLO-205 Cells (Colon Adenocarcinoma Cell, CCL-222) COLO-205 Cell Line Description and Applications COLO-205 Cells www.Altogen.com The COLO-205 cell line was established from the cell line CV-1, a cell line established from the kidney cells of an African Green monkey (Cercopithecus aethiops), that was transformed with a mutant SV-40 genome. This SV-40 genome produces large T antigens. The COLO-205 cell line is characterized as fibroblast-like cells that grow as a monolayer. Researchers use the COLO-205 cell line to study the SV-40 monkey virus as well as to transfect cells. As a tool for transfection, the COLO-205 cell line has applications for the production of recombinant proteins for biochemistry as well as molecular and cellular biology. Research indicates that COLO-205 can completely allow for lytic growth of SV-40. 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 COLO-205 Cells (Colon Adenocarcinoma Cell, CCL-222) COLO-205 Transfection Protocol 1. Plate 10,000 - 15,000 COLO-205 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 COLO-205 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 COLO-205 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 COLO-205 Cells (Colon Adenocarcinoma Cell, CCL-222) 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 COLO-205 Cells (Colon Adenocarcinoma Cell, CCL-222) COLO-205 Transfection Kit Product Details • A nanoparticle-based liposome formulation • 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 COLO-205 Cells (Colon Adenocarcinoma Cell, CCL-222) Data Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into COLO-205 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 COLO-205 Cells (Colon Adenocarcinoma Cell, CCL-222) Data Figure 2. Protein expression of Lamin A/C in COLO-205 cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into COLO-205 cells following Altogen Biosystems transfection protocol. At 72 hours post-transfection 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 COLO-205 Cells (Colon Adenocarcinoma Cell, CCL-222) COLO-205 Transfection Kit Benefits • Pre-optimized transfection protocol for COLO-205 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 COLO-205 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 COLO-205 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]