Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) Altogen Biosystems offers the IMR-90 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The IMR-90 Transfection Reagent is an advanced formulation of a lipid-based reagent, and it has been developed to provide high transfection efficiency with the IMR-90 cell line. This cell line is a good host for a variety of research related to molecular and cellular biology. When cultured in vitro, IMR-90 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 IMR-90 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 IMR-90 cells atcc.org Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) IMR-90 Cell Line Description and Applications The IMR-90 cell line originates from lung tissue from a 16 week old female Caucasian fetus derived by W. W. Nichols and associates. The cell line is a human diploid strain and displays fibroblast morphology. This cell line has applications as a model of fibroblasts, but it also demonstrates some characteristics of differentiated smooth muscle cells. Research indicates that the IMR-90 cell line can reach approximately 58 population doublings prior to senescence. IMR-90 cells have a rate of contraction faster than other established cell lines when cultivated on a rubbery silicon surface. High Density IMR-90 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

IMR-90 Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) IMR-90 Transfection Protocol 1. Plate 10,000 - 15,000 IMR-90 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 IMR-90 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 IMR-90 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 > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) 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

IMR-90 Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) IMR-90 Transfection Kit Product Details Two-component formulation enhances lipid-mediated transfection efficiency 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

Data Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in the IMR-90 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).

Data Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) Data Figure 2. Protein expression of Cyclophilin B in IMR-90 cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into IMR-90 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.

IMR-90 Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186) IMR-90 Transfection Kit Benefits Pre-optimized transfection protocol for IMR-90 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 IMR-90 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 IMR-90 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