Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163) Altogen Biosystems offers the BALB/3T3 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The BALB/3T3 Transfection Reagent is a nanoparticle-based liposome formulation reagent that enhances lipid-mediated transfection, and it has been developed to provide high transfection efficiency with BALB/3T3 cells. This cell line is an excellent host for transfection and the assessment of cellular functionality. When cultured in vitro, BALB/3T3 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 BALB/3T3 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 BALB/3T3 Cells Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163) BALB/3T3 Cell Line Description and Applications The BALB cell line is also known as the 3T3 cell line (clone A31). The cell line originated from Mus musculus embryos and was established in 1968. For optimal BALB cell growth, care must be given to environment as the BALB cell line is susceptible to environmental factors. The BALB cell line is particularly useful for studying mammal embryonic stem cells such as stem cells. Furthermore, this BALB/3T3 cell line can serve as a transfection host in a spectrum of biology research. High Density BALB/3T3 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

BALB/3T3Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163) BALB/3T3Transfection Protocol 1. Plate 10,000 - 15,000 BALB/3T3 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 BALB/3T3cells (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 BALB/3T3cells 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 > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163) 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

BALB/3T3Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163) BALB/3T3Transfection 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 trust4 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

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163) Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in BALB/3T3cell line 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 > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163) Data Figure 2. Protein expression of Lamin A/C in BALB/3T3 cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into BALB/3T3 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.

BALB/3T3Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163) BALB/3T3Transfection Kit Benefits Pre-optimized transfection protocol for BALB/3T3 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 BALB/3T3 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 BALB/3T3 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