Products > AsPC-1 Transfection Reagent (Pancreatic Beta Cells)

1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products >AsPC-1 Transfection Reagent (Pancreatic Beta Cells) Altogen Biosystems offers the AsPC-1 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. AsPC-1 Transfection Reagent is an advanced formulation of a nanoparticle-based liposome reagent, and it has been developed to provide high transfection efficiency with AsPC-1 and other pancreatic cancer cell lines. This cell line is an excellent host for studying pancreatic cancer and human infections related to the pancreas. When cultured in vitro, AsPC-1 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 AsPC-1 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 >AsPC-1 Transfection Reagent (Pancreatic Beta Cells) AsPC-1Cell Line Description and Applications The AsPC-1 cell line was established from a 62-year-old Caucasian woman with metastatic pancreatic adenocarcinoma. It was reestablished through a nude mouse xenograft with cells from the ascites, and the reestablished epithelial cells retained the original morphology. The AsPC-1 cells can be used to study therapeutics for pancreatic cancer. This is a crucial area of research as the five-year survival rate of pancreatic cancer is less than 1%. Recent research has shown sanguinarine, a plant alkaloid, and deuterium oxide (D2O), or heavy water, induce apoptosis of AsPC-1. AsPC-1 cellswww.clsgmbh.de 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 >AsPC-1 Transfection Reagent (Pancreatic Beta Cells) AsPC-1Transfection Protocol 1. Plate 10,000 - 15,000 AsPC-1 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 AsPC-1cells (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 AsPC-1cells 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 >AsPC-1 Transfection Reagent (Pancreatic Beta 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 >AsPC-1 Transfection Reagent (Pancreatic Beta Cells) AsPC-1Transfection Kit Product Details • A nanoparticle-based liposome formulation • Optimized easy-to-use transfection protocol provided for transfection of siRNA, DNA, mRNA, and microRNA • Kit includes Transfection Enhancer reagent and recommended transfection protocol • High transfection efficacy in the presence of serum • Expand your RNAi application with a reagent optimized for delivery of both siRNA and plasmid • Reproducible transfection results • Works well for standard reverse transfection and high-throughput applications • 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 >AsPC-1 Transfection Reagent (Pancreatic Beta Cells) Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in the AsPC-1 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 >AsPC-1 Transfection Reagent (Pancreatic Beta Cells) Data Figure 2. Protein expression of Cyclophilin B in AsPC-1 cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into AsPC-1 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 >AsPC-1 Transfection Reagent (Pancreatic Beta Cells) AsPC-1Transfection Kit Benefits • Pre-optimized transfection protocol for AsPC-1 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 AsPC-1 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 AsPC-1 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