Products > MRC-5 Transfection Reagent (Diploid Fibroblast Cells)

1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > MRC-5 Transfection Reagent (Diploid Fibroblast Cells) Altogen Biosystems offers the MRC-5 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The MRC-5 Transfection Reagent is an advanced nanoparticle-based liposome formulation, and it has been developed to provide high transfection efficiency with the MRC-5 cell line. This cell line is a good host for virology, cytotoxicity, and transfection. When cultured in vitro, MRC-5 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 MRC-5 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 > MRC-5 Transfection Reagent (Diploid Fibroblast Cells) MRC-5 Cell Line Description and Applications The MRC-5 cell line originates from the healthy pulmonary tissue of an aborted 14-week-old male fetus in September of 1966 by J.P. Jacobs. This cell line is a normal diploid cell line and as a result, has a modal chromosome number of 46. The MRC-5 cell line displays adherent growth and has the capacity for approximately 42-46 population doublings prior to senescence. The cell line can support a wide range of viruses including herpes simplex virus, poliovirus, rhinovirus and varicella-zoster virus. As a result, the MRC-5 cell line has several applications for research in the field of virology, but also has applications for research related to vaccines, transfection and cytotoxicity assessment. High Density MRC-5 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

3. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > MRC-5 Transfection Reagent (Diploid Fibroblast Cells) MRC-5 Transfection Protocol 1. Plate 10,000 - 15,000 MRC-5 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 MRC-5 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 MRC-5 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

4. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > MRC-5 Transfection Reagent (Diploid Fibroblast 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 > MRC-5 Transfection Reagent (Diploid Fibroblast Cells) MRC-5 Transfection Kit Product Details • A nanoparticle-based liposome formulation • Transfection protocols provided for transfection of proteins, DNA, mRNA, siRNA, shRNA and microRNA • Transfection Enhancer reagent provided with the kit • Produce higher level of recombinant protein expression with minimal disruption of normal cell function • Generate physiologically relevant data you can trust • Effective for plasmid DNA/siRNA co-transfection • Easy-to-use transfection protocol with reproducible results • Low cytotoxicity • 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 > MRC-5 Transfection Reagent (Diploid Fibroblast Cells) Data Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into MRC-5 cells following the recommended protocol. At 48 hours post-transfection the cells were analyzed by qRT-PCR for Lamin A/C gene expression levels. 18S rRNA levels were used to normalize the Lamin A/C data. Values are normalized to untreated sample. Data are means ± SD (n=4).

7. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > MRC-5 Transfection Reagent (Diploid Fibroblast Cells) Data Figure 2. Protein expression of Lamin A/C in MRC-5 cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into MRC-5 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 > MRC-5 Transfection Reagent (Diploid Fibroblast Cells) MRC-5 Transfection Kit Benefits • Pre-optimized transfection protocol for MRC-5 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 MRC-5 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 MRC-5 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