1. Altogen Biosystems offers the Transfection Reagent for PANC-1 Cells Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The PANC-1 Transfection Reagent is a proprietary cationic lipids formulation that enhances lipid mediated transfection, and it has been developed to provide high transfection efficiency with PANC- 1 cells. This cell line has applications for research related to pancreatic cancer. When cultured in vitro, PANC-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 PANC-1 Transfection Kit at www.Altogen.com Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)

2. 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 PANC-1 Cells Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer) The PANC-1 cell line is an abbreviation for “pancreas,” and originates from pancreatic duct tissue of a 56-year old Caucasian male patient suffering from epithelioid carcinoma. The PANC-1 cell line has a population doubling time of 52 hours and a chromosomal modal number of 63. The cell line can be used as an in vitro model in tumorigenicity studies related to non-endocrine pancreatic cancer. L-asparagine inhibits growth of PANC-1 cells. FGF2 stimulates the cell line to differentiate into islet like clusters that produce hormones. The PANC-1 cell line is negative for estrogen receptor. The cell line has particular applications for research related to pancreas cancer, but also serves an excellent transfection host.

3. PANC-1 Transfection Protocol 1. Plate 10,000 - 15,000 PANC-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 PANC-1 cells (from step 2) 7. Incubate cells at 37ºC in a humidified CO 2 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 PANC-1 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 Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)

4. 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 Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)

5. PANC-1 Transfection Kit Product Details Proprietary 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 Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)

6. Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in PANC-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). Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)

7. Data Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Figure 2. Protein expression of Cyclophilin B in PANC-1 cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into PANC-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. Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)

8. Pre-optimized transfection protocol for PANC-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 PANC-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 PANC-1 cell lines Bio-degradable after endocytosis Used for preclinical research worldwide PANC-1 Transfection Kit Benefits 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 > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)