Transfection Reagent for Chromaffin Cells

1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products >Transfection Reagent for Chromaffin Cells Altogen Biosystems offers the Transfection Reagent for Chromaffin Cells Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The Chromaffin Transfection Reagent is an advanced nanoparticle-based liposome formulated reagent that enhances lipid-mediated transfection, and it has been developed to provide high transfection efficiency with Chromaffin cells. This cell line has applications for research in the fields of cellular and molecular biology. When cultured in vitro, Chromaffin 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 Chromaffin 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 >Transfection Reagent for Chromaffin Cells Chromaffin Cell Line Description and Applications The Chromaffin cell line was established from chromaffin cell cultures that were also named Medullary Chromaffin cells. The name Chromaffin is a portmanteau of “chromium” and “affinity” because the cell line is easily visible with a chromatin stain. Chromaffin cells are a type of neuroendocrine cells. Neuroendocrine cells are located in the sympathetic nervous system and are responsible for several processes including the “fight or flight” response. Chromaffin cells have implications for a variety of areas in research in the fields of molecular and cellular biology. Chromaffin Cells www.histology-world.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 >Transfection Reagent for Chromaffin Cells Chromaffin Transfection Protocol 1. Plate 10,000 - 15,000 Chromaffin 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 Chromaffin 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 Chromaffin 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 >Transfection Reagent for Chromaffin 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 >Transfection Reagent for Chromaffin Cells Chromaffin Transfection Kit Product Details • A biodegradable polymer-based transfection agent – once inside the cell, the polymer degrades into smaller less toxic components reducing cell toxicity, facilitating release of the transgene, and improving 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 >Transfection Reagent for Chromaffin Cells Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in Chromaffin 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=3).

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