Products > Beta-TC-6 Transfection Reagent (Pancreas Tumor, CRL-11506)

1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Beta-TC-6 Transfection Reagent (Pancreas Tumor, CRL-11506) Altogen Biosystems offers the Beta-TC-6Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The Beta-TC-6 Transfection Reagent is a two-component lipid-mediated formulation, and it has been developed to provide high transfection efficiency with the Beta-TC-6 cell line. This cell line is particularly useful for researching diabetes. When cultured in vitro, Beta-TC-6 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 Beta-TC-6 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 > Beta-TC-6 Transfection Reagent (Pancreas Tumor, CRL-11506) Beta-TC-6 Cell Line Description and Applications The Beta-TC-6 cell line originates from tumorigenic pancreatic tissue derived from a transgenic mouse or Mus musculus with insulinoma. These cells produce glucagon, somatostatin, and insulin. Insulin, in particular, is abundantly produced and secreted mainly as a response to glucose. The Beta-TC-6 display beta cell and epithelial cell morphology. They are also transgenic in regards to the SV40 large T antigen because the mouse carried a construct made up of the SV40 early region. High Density Beta-TC-6 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 > Beta-TC-6 Transfection Reagent (Pancreas Tumor, CRL-11506) Beta-TC-6 Transfection Protocol 1. Plate 10,000 - 15,000 Beta-TC-6 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 Beta-TC-6 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 Beta-TC-6 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 > Beta-TC-6 Transfection Reagent (Pancreas Tumor, CRL-11506) 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 > Beta-TC-6 Transfection Reagent (Pancreas Tumor, CRL-11506) Beta-TC-6 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

6. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Beta-TC-6 Transfection Reagent (Pancreas Tumor, CRL-11506) Data Figure 1. GAPDH mRNA levels were quantified using real-time RT-PCR in the Beta-TC6 cells transfected with siRNAs targeting GAPDH or non-silencing siRNA. Forty-eight hours post-transfection, the cells were harvested and analyzed by real-time RT-PCR for GAPDH mRNA expression levels. Data were normalized against the 18S rRNA signal. Control samples were either mock-transfected or untreated. Values are normalized to untreated sample. Data are means ± SD (n=5).

7. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Beta-TC-6 Transfection Reagent (Pancreas Tumor, CRL-11506) Data Figure 2. Protein expression of GAPDH in Beta-TC-6 cells. DNA plasmid expressing GAPDH or siRNA targeting GAPDH were transfected into Beta-TC-6 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 > Beta-TC-6 Transfection Reagent (Pancreas Tumor, CRL-11506) Beta-TC-6 Transfection Kit Benefits • Pre-optimized transfection protocol for Beta-TC-6 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 Beta-TC-6 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 Beta-TC-6 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