Products > HeLa Transfection Reagent (Cervix Adenocarcinoma Cells)

1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HeLa Transfection Reagent (Cervix Adenocarcinoma Cells) Altogen Biosystems offers the HeLa Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The HeLa Transfection Reagent is a liposome formulationreagent, and it has been developed to provide high transfection efficiency with the HeLa cell line. This cell line is a good host for a wide variety of research in cell and molecular biology. When cultured in vitro, HeLa 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 HeLa 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 > HeLa Transfection Reagent (Cervix Adenocarcinoma Cells) HeLa Cell Line Description and Applications The HeLa cell line originates from the cervical cancer cells of Henrietta Lacks, a 31-year old female patient, in 1951. The HeLa cell line is remarkable in that it is the first human cell line able to survive in vitro indefinitely. The cells were taken from a tissue sample from the adenocarcinoma, and HeLa cells have a modal chromosome number of 82. HeLa cells have an active type of telomerase during cell division, and as a result, the cell line proliferates extremely quickly in comparison to other cell lines. Jonas Salk used HeLa cells to test the polio vaccine, and HeLa cells have been an invaluable tool in a vast array of research from cancer to gene mapping to AIDS and studying the toxic effects of radiation. The HeLa cell line contains human papillomavirus sequences and tests positive for lyso-PC and keratin. HeLa cells Atcc.com 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 > HeLa Transfection Reagent (Cervix Adenocarcinoma Cells) HeLa Transfection Protocol 1. Plate 10,000 - 15,000 HeLa 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 HeLa 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 HeLa 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 > HeLa Transfection Reagent (Cervix Adenocarcinoma Cells) 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 > HeLa Transfection Reagent (Cervix Adenocarcinoma Cells) HeLa Transfection Kit Product Details • Proprietary cationic lipids 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  techserv@altogen.com

6. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HeLa Transfection Reagent (Cervix Adenocarcinoma Cells) Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in HeLa 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=3).

7. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HeLa Transfection Reagent (Cervix Adenocarcinoma Cells) Data Figure 2. Protein expression of Cyclophilin B in HeLa cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into HeLa 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 > HeLa Transfection Reagent (Cervix Adenocarcinoma Cells) HeLa Transfection Kit Benefits • Pre-optimized transfection protocol for HeLa 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 HeLa 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 HeLa 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