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Altogen Biosystems  848 Rainbow Blvd #823  L as Vegas  N V  8 9107  U SA

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products. Products > HCT-116 Transfection Reagent (Colon Carcinoma).

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Altogen Biosystems  848 Rainbow Blvd #823  L as Vegas  N V  8 9107  U SA

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  1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HCT-116 Transfection Reagent (Colon Carcinoma) Altogen Biosystems offers the HCT-116 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The HCT-116 Transfection Reagent is an advanced formulation of a biodegradable polymer-basedreagent, and it has been developed to provide high transfection efficiency with the HCT-116 cell line. This cell line is a good host for studying kidney cell function. When cultured in vitro, HCT-116 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 HCT-116 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

  2. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HCT-116 Transfection Reagent (Colon Carcinoma) HCT-116 Cell Line Description and Applications The HCT-116 cell line originates from epithelial tumorigenic tissue taken from the colon of an adult male with a colorectal carcinoma. The cells are positive for transforming growth factor and keratin by immunoperoxidase. The HCT-116 cell line has a mutation in codon 13 of the ras proto-oncogene and as a result, the cell line has applications for PCR studies related to the colon. This cell line is used to study therapeutics associated with colon cancer. High Density HCT-116 cells www.atcc.org Altogen Biosystems 848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone 702 349 6103  Fax 702 989-0841  email  techserv@altogen.com

  3. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HCT-116 Transfection Reagent (Colon Carcinoma) HCT-116 Transfection Protocol 1. Plate 10,000 - 15,000 HCT-116 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 HCT-116 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 HCT-116 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

  4. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HCT-116 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

  5. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HCT-116 Transfection Reagent (Colon Carcinoma) HCT-116 Transfection Kit Product Details • Proprietary cationic lipids formulation • Optimized for intracellular delivery of plasmid DNA, siRNA, microRNA, and mRNA • High transfection efficiency of both siRNA and plasmid DNA without compromising cell viability • Achieve robust siRNA uptake for dependable gene silencing • Effective transfection under conditions of up to 40% serum • Transfection kit includes Transfection Enhancer reagent • Gentle enough to be used for single cell analysis • 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

  6. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HCT-116 Transfection Reagent (Colon Carcinoma) Data Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into HCT-116 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).

  7. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HCT-116 Transfection Reagent (Colon Carcinoma) Data Figure 2. Protein expression of Lamin A/C in HCT-116 cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into HCT-116 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.

  8. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HCT-116 Transfection Reagent (Colon Carcinoma) HCT-116 Transfection Kit Benefits • Pre-optimized transfection protocol for HCT-116 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 HCT-116 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 HCT-116 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

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