Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PGHAM Transfection Reagent (Hamster Pancreatic Carcinoma) Altogen Biosystems offers the PGHAM Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. PGHAM Transfection Reagent is a two component formulation that enhances lipid-mediated transfection efficiency, and it is developed to provide high transfection efficiency with PGHAM and related cell lines. This cell line is a good host for studying pancreatic cancer. Other applications include drug discovery, gene expression studies, molecular and cell biology research applications. Purchase PGHAM 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

PGHAM Cell Line Description and Applications Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PGHAM Transfection Reagent (Hamster Pancreatic Carcinoma) PGHAM Cell Line Description and Applications The PGHAM cell line originates from a pancreatic tumor in a Syrian hamster (Mesocricetus auratus) induced by N-nitrosobis(2-oxopropyl) (BOP). 6-8 weeks after transplantation, the recipient hamsters were called and after 8 subcutaneous transplantations, the tumor was extracted. The tumor displayed characteristics similar to a pancreatic ductal adenocarcinoma. One of the most common K-ras mutations in pancreatic cancers is a point mutation at codon 12 and PGHAM has this mutation. The PGHAM cell line is known for its rapid growth and high rate of metastasis, and it is particularly useful for studying therapeutics related to pancreatic cancer. PGHAM Cell Line cancerres.aacrjournals.org Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

PGHAM Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PGHAM Transfection Reagent (Hamster Pancreatic Carcinoma) PGHAM Transfection Protocol Plate 10,000 - 15,000 PGHAM cells per well in 0.5 ml of complete growth medium 12–24 hours prior to transfection Wash with 1xPBS and add 0.5 ml of fresh growth medium 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 Incubate transfection complexes at RT for 15 - 30 minutes 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 Add prepared transfection complexes to 0.5 ml of complete growth medium with PGHAM cells (from step 2) Incubate cells at 37ºC in a humidified CO2 incubator 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 PGHAM 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

General Lipoplex-mediated Transfection Mechanism of Action Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PGHAM Transfection Reagent (Hamster Pancreatic Carcinoma) 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

PGHAM Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PGHAM Transfection Reagent (Hamster Pancreatic Carcinoma) PGHAM Transfection Kit Product Details Two-component formulation enhances lipid-mediated transfection efficiency 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

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

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PGHAM Transfection Reagent (Hamster Pancreatic Carcinoma) Data Figure 2. Protein expression of Cyclophilin B in PGHAM cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into PGHAM 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.

PGHAM Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PGHAM Transfection Reagent (Hamster Pancreatic Carcinoma) PGHAM Transfection Kit Benefits Pre-optimized transfection protocol for PGHAM 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 PGHAM 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 PGHAM 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