Products > HUVEC Transfection Reagent (Human Umbilical Vein Endothelial Cells)

1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > HUVEC Transfection Reagent (Human Umbilical Vein Endothelial Cells) Altogen Biosystems offers the HUVECTransfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The HUVEC Transfection Reagent is an advanced formulation of a lipid-basedreagent, and it has been developed to provide high transfection efficiency with the HUVEC cell line. This cell line is a good host for studying angiogenesis and cancer therapeutics. When cultured in vitro, HUVEC 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 HUVEC 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 > HUVEC Transfection Reagent (Human Umbilical Vein Endothelial Cells) HUVEC Cell Line Description and Applications The HUVEC or human umbilical vein endothelial cell line originates, as the name suggests, from a human umbilical vein and is an extremely useful tool for research relating to the umbilical cord. Research indicates that HUVEC cells may participate in angiogenesis, the process by which blood vessels form from pre-existing vessels. Angiogenesis can also literally feed cancerous growths with nutrients. Some cancer therapy research is based upon the targeting of angiogenesis. The HUVEC cell line also provides a tool for research related to macromolecule transport, blood coagulation, and fibrinolysis. HUVEC cells 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 > HUVEC Transfection Reagent (Human Umbilical Vein Endothelial Cells) HUVEC Transfection Protocol 1. Plate 10,000 - 15,000 HUVEC 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 HUVEC 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 HUVEC 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 > HUVEC Transfection Reagent (Human Umbilical Vein Endothelial 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 > HUVEC Transfection Reagent (Human Umbilical Vein Endothelial Cells) HUVEC Transfection Kit Product Details • 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 > HUVEC Transfection Reagent (Human Umbilical Vein Endothelial Cells) Data Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the HUVEC 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 GAPDH 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=3).

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