DI-TNC1 Transfection Reagent (Rat Brain Astrocytes)

1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > DI-TNC1 Transfection Reagent (Rat Brain Astrocytes) Altogen Biosystems offers the DI-TNC1 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The DI-TNC1 Transfection Reagent is an advanced formulation of a nanoparticle-based liposome formulation, and it has been developed to provide high transfection efficiency with the DI-TNC1 cell line. This cell line is a good host for studying astrocytes, glia and neurons. When cultured in vitro, DI-TNC1 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 DI-TNC1 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 > DI-TNC1 Transfection Reagent (Rat Brain Astrocytes) The D1-TNC1 cell line originates from the primary cultures of Type 1 astrocytes taken from the brain diencephalon or interbrain tissue of 1-day old rats. After initial plating, the cultures were transfected with a DNA construct. The DNA construct contained the oncogenic region of SV40 under the transcriptional control of the human GFAP promoter (pGFA-Tt). The DI-TNC1 cell line has characteristics of Type 1 astrocytes such as GFAP immunoreactivity, but also produces transferrin in lesser amounts. Over 95% of D1-TNC1 have been shown to contain the SV40 T antigen. This cell line can be used to study neurons, astrocytes and glia. DI-TNC1 cells Lonza.com 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 > DI-TNC1 Transfection Reagent (Rat Brain Astrocytes) DI-TNC1 Transfection Protocol 1. Plate 10,000 - 15,000 DI-TNC1 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 DI-TNC1 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 DI-TNC1 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 > DI-TNC1 Transfection Reagent (Rat Brain Astrocytes) 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 > DI-TNC1 Transfection Reagent (Rat Brain Astrocytes) DI-TNC1 Transfection Kit Product Details • A nanoparticle-based liposome 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

6. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > DI-TNC1 Transfection Reagent (Rat Brain Astrocytes) Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in the DI-TNC1 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).

7. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > DI-TNC1 Transfection Reagent (Rat Brain Astrocytes) Data Figure 2. Protein expression of Cyclophilin B in DI-TNC1 cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into DI-TNC1 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 > DI-TNC1 Transfection Reagent (Rat Brain Astrocytes) DI-TNC1 Transfection Kit Benefits • Pre-optimized transfection protocol for DI-TNC1 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 DI-TNC1 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 DI-TNC1 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