Products > CT26.WT Transfection Reagent (Colon Carcinoma) Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > CT26.WT Transfection Reagent (Colon Carcinoma) Altogen Biosystems offers the CT26.WT Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. CT26.WT Transfection Reagent is an advanced formulation of a cationic lipid-based reagent, and it is developed to provide high transfection efficiency with CT26.WT and other colon cancer cell lines. This cell line is a good host for studying colon cancer and immunotherapies. When cultured in vitro, CT26.WT 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 CT26.WT 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  techserv@altogen.com

High Density Image of CT26.WT Cells Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > CT26.WT Transfection Reagent (Colon Carcinoma) CT26.WT Cell Line Description and Applications The C26.WT originates from colon carcinoma cells and was stably transduced. The CT26.WT cell line was cloned with the retrovirus vector LXSN that contained the lacZ gene. This cell line is undifferentiated, tumorigenic and displays fibroblast morphology. The cell line has applications for discovering therapeutics to combat colon cancer, a prevalent disease as well as studies related to immunotherapy protocols and responses. In fact, the CT26.WT cell line is one of the most frequently employed cell lines for drug development. High Density Image of CT26.WT Cells Imanislife.com Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

CT26.WT Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > CT26.WT Transfection Reagent (Colon Carcinoma) CT26.WT Transfection Protocol 1. Plate 10,000 - 15,000 CT26.WT 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 CT26.WT 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 CT26.WT 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  techserv@altogen.com

General Lipoplex-mediated Transfection Mechanism of Action Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > CT26.WT Transfection Reagent (Colon Carcinoma) 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  techserv@altogen.com

CT26.WT Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > CT26.WT Transfection Reagent (Colon Carcinoma) CT26.WT Transfection Kit Product Details Two component formulation enhances lipid mediated transfection efficiency Optimized easy-to-use transfection protocol provided for transfection of siRNA, DNA, mRNA, and microRNA Kit includes Transfection Enhancer reagent and recommended transfection protocol High transfection efficacy in the presence of serum Expand your RNAi application with a reagent optimized for delivery of both siRNA and plasmid Reproducible transfection results Works well for standard reverse transfection and high-throughput applications 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  techserv@altogen.com

Data Products > CT26.WT Transfection Reagent (Colon Carcinoma) Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > CT26.WT Transfection Reagent (Colon Carcinoma) Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in CT26WT cells transfected by Cyclophilin B siRNA or non-silencing siRNA control following the recommended transfection protocol. Cyclophilin mRNA expression levels were measured 48 hours post-transfection. 18S rRNA levels were used to normalize the Cyclophilin B data. Values are normalized to untreated sample. Data are presented as means ± SD (n=6).

Data Products > CT26.WT Transfection Reagent (Colon Carcinoma) Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > CT26.WT Transfection Reagent (Colon Carcinoma) Data Figure 2. Protein expression of Cyclophilin B in CT26.WT cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into CT26.WT 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.

CT26.WT Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > CT26.WT Transfection Reagent (Colon Carcinoma) CT26.WT Transfection Kit Benefits Pre-optimized transfection protocol for CT26.WT 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 CT26.WT 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 CT26.WT 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  techserv@altogen.com