Products > MIA PaCa-2 Transfection Reagent (Pancreatic Carcinoma) Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > MIA PaCa-2 Transfection Reagent (Pancreatic Carcinoma) Altogen Biosystems offers the MIA PaCa-2 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The MIA PaCa-2 Transfection Reagent is an advanced formulation of a lipid-based reagent, and it has been developed to provide high transfection efficiency with the MIA PaCa-2 cell line. This cell line is a good host for medical research related to pancreatic adenocarcinoma. When cultured in vitro, MIA PaCa-2 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 MIA PaCa-2 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

High Density MIA PaCa-2 cells Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > MIA PaCa-2 Transfection Reagent (Pancreatic Carcinoma) MIA PaCa-2 Cell Line Description and Applications The MIA PaCa-2 cell line originates from tissue derived from a pancreatic tumor taken from a 65-year old Caucasian male by A. Yunis et al. in 1975. This cell line has an estimated population doubling time of approximately 40 hours. The MIA PaCa-2 cell line is hypotriploid and displays epithelial morphology and rounded, floating cells. The modal chromosome number for this cell line is 61, and a few normal chromosomes are absent. The MIA PaCa-2 cell line produces CSF-1 as well as plasminogen activators. High Density MIA PaCa-2 cells atcc.org Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

MIA PaCa-2 Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > MIA PaCa-2 Transfection Reagent (Pancreatic Carcinoma) MIA PaCa-2 Transfection Protocol 1. Plate 10,000 - 15,000 MIA PaCa-2 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 MIA PaCa-2 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 MIA PaCa-2 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 > MIA PaCa-2 Transfection Reagent (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

MIA PaCa-2 Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > MIA PaCa-2 Transfection Reagent (Pancreatic Carcinoma) MIA PaCa-2 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

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > MIA PaCa-2 Transfection Reagent (Pancreatic Carcinoma) Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in the MIA Paca-2 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 > MIA PaCa-2 Transfection Reagent (Pancreatic Carcinoma) Data Figure 2. Protein expression of Cyclophilin B in MIA PaCa-2 cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into MIA PaCa-2 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.

MIA PaCa-2 Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > MIA PaCa-2 Transfection Reagent (Pancreatic Carcinoma) MIA PaCa-2 Transfection Kit Benefits Pre-optimized transfection protocol for MIA PaCa-2 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 MIA PaCa-2 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 MIA PaCa-2 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