Products > CHO-K1 Transfection Reagent (Ovarian Adenocarcinoma)

1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > CHO-K1 Transfection Reagent (Ovarian Adenocarcinoma) Altogen Biosystems offers the CHO-K1Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The CHO-K1 Transfection Reagent is an advanced formulation of a liposome-based lipid reagent, and it has been developed to provide high transfection efficiency with the CHO-K1 cell line. This cell line is a good transfection host and a tool for research related to ovarian adenocarcinoma. When cultured in vitro, CHO-K1 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 CHO-K1 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 > CHO-K1 Transfection Reagent (Ovarian Adenocarcinoma) CHO-K1 Cell Line Description and Applications The CHO-K1 cell line originates from a subclone of the CHO cell line, a cell line established in 1957 and derived from the ovarian tissue of an adult Chinese hamster (Cricetulus griseus). The CHO-K1 cell line requires proline for cell growth but nonetheless makes an excellent transfection host. The cell line is resistant to poliovirus 2, Modoc virus and Button Willow virus. The CHO-K1 cell line was mutagenized with ethyl methane sulfonate and this generated a cell line lacking DHFR. High Density CHO-K1 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

3. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > CHO-K1 Transfection Reagent (Ovarian Adenocarcinoma) CHO-K1 Transfection Protocol 1. Plate 10,000 - 15,000 CHO-K1 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 CHO-K1 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 CHO-K1 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 > CHO-K1 Transfection Reagent (Ovarian Adenocarcinoma) 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 > CHO-K1 Transfection Reagent (Ovarian Adenocarcinoma) CHO-K1 Transfection Kit Product Details • A liposome-based lipid 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 > CHO-K1 Transfection Reagent (Ovarian Adenocarcinoma) Data Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into CHO-K1 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 > CHO-K1 Transfection Reagent (Ovarian Adenocarcinoma) Data Figure 2. Protein expression of Lamin A/C in CHO-K1 cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into CHO-K1 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 > CHO-K1 Transfection Reagent (Ovarian Adenocarcinoma) CHO-K1 Transfection Kit Benefits • Pre-optimized transfection protocol for CHO-K1 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 CHO-K1 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 CHO-K1 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