Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Caco-2 Cells Transfection Reagent (Colon Adenocarcinoma Cell, HTB-37) Altogen Biosystems offers the Caco-2 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The Caco-2 Transfection Reagent is a biodegradable polymer based transfection reagent that enhances transfection, and it has been developed to provide high transfection efficiency with Caco-2 cells. This cell line is a good host for transfection and the assessment of cellular functionality. It is also an excellent host for the study of colon cancers. When cultured in vitro, Caco-2 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 Caco-2 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 Caco-2 Cells Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Caco-2 Cells Transfection Reagent (Colon Adenocarcinoma Cell, HTB-37) Caco-2 Cell Line Description and Applications The Caco-2 cell line is a heterogeneous epithelial colorectal adenocarcinoma cell line that originates from the cancerous colon cells of a 72-year old Caucasian patient. The cell line was isolated by Dr. Jorgen Fogh via the Sloan-Kettering Institute. The cell has doubles approximately 62 hours. The Caco-2 cell line has the potential to be used as a tool for researching a variety of colon cancers and is vulnerable to human immunodeficiency virus 1. The cell line can differentiate and resemble small intestine enterocytes and also expresses several enzymes and transporters. High Density Caco-2 Cells 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

Caco-2 Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Caco-2 Cells Transfection Reagent (Colon Adenocarcinoma Cell, HTB-37) Caco-2 Transfection Protocol 1. Plate 10,000 - 15,000 Caco-2 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 Caco-2 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 Caco-2 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 > Caco-2 Cells Transfection Reagent (Colon Adenocarcinoma Cell, HTB-37) 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

Caco-2 Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Caco-2 Cells Transfection Reagent (Colon Adenocarcinoma Cell, HTB-37) Caco-2 Transfection Kit Product Details A biodegradable polymer based transfection agent – once inside the cell, the polymer degrades into smaller less toxic components reducing cell toxicity, facilitating release of the transgene, and improving 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 trust4 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  techserv@altogen.com

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Caco-2 Cells Transfection Reagent (Colon Adenocarcinoma Cell, HTB-37) Data Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the Caco-2 cells transfected with siRNAs targeting GAPD or non-silencing siRNA. Forty-eight hours post-transfection, the cells were harvested and analyzed by real-time RT-PCR for GAPD mRNA expression levels. Data were normalized against the 18S rRNA signal. Control samples were either mock-transfected or untreated. Values are normalized to untreated sample. Data are means ± SD (n=5).

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Caco-2 Cells Transfection Reagent (Colon Adenocarcinoma Cell, HTB-37) Data Figure 2. Protein expression of GAPDH in Caco-2 cells. DNA plasmid expressing GAPDH or siRNA targeting GAPDH were transfected into Caco-2 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.

Caco-2 Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Caco-2 Cells Transfection Reagent (Colon Adenocarcinoma Cell, HTB-37) Caco-2 Transfection Kit Benefits Pre-optimized transfection protocol for Caco-2 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 Caco-2 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 Caco-2 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