Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for MDA-MB Cells (Breast Adenocarcinoma Cells, HTB-26) Altogen Biosystems offers the MDA-MB Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The MDA-MB Transfection Reagent two component formulation enhances lipid mediated transfection efficiency, and it has been developed to provide high transfection efficiency with the MDA-MB-231, MDA-MB-435, MDA-MB-468 cell lines. This cell line is a good host for studying breast cancer proliferation. When cultured in vitro, MDA-MB 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 MDA-MB 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

MDA-MB Cell Line Description and Applications Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for MDA-MB Cells (Breast Adenocarcinoma Cells, HTB-26) MDA-MB Cell Line Description and Applications The MDA-MB category includes several cell lines such as MDA-MB-231, MDA-MB-435 and MDA-MB-468, many of which possess varying characteristics. In fact, some MDA-MB cell lines form colonies in a semisolid medium, but are not tumorigenic in immunosuppressed mice. The MDA-MB-453 cells in particular originated in 1976 from an effusion of a 48-year old Caucasian female. The patient suffered from metastatic carcinoma of the breast, and it involved the nodes and brain as well as both pericardial and pleural cavities. MDA-MB-453 cells overexpress FGF receptors and are round, epithelial-like, loosely attached monolayers. Research shows that MDA-MB-453 cells have an active glycerol 3-phosphate shuttle, and that that these cells can provide an in vitro model for androgen regulation of gene expression as they produce high levels of functional androgen receptors. Via research with this cell line, scientists have discovered that G-protein coupled inwardly in response to potassium (GIRK) is involved in cellular signaling in breast cancer cells. MDA-MB 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

MDA-MB Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for MDA-MB Cells (Breast Adenocarcinoma Cells, HTB-26) MDA-MB Transfection Protocol 1. Plate 10,000 - 15,000 MDA-MB 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 MDA-MB 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 MDA-MB 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

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 MDA-MB Cells (Breast Adenocarcinoma Cells, HTB-26) 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

MDA-MB Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for MDA-MB Cells (Breast Adenocarcinoma Cells, HTB-26) MDA-MB Transfection Kit Product Details Optimized for MDA-MB-231, MDA-MB-435, MDA-MB-468 cell lines Two component formulation enhances lipid mediated transfection efficiency Optimized easy-to-use transfection protocol provided for transfection of siRNA, DNA, mRNA, and microRNA Kit includes Transfection Enhancer reagent and recommended transfection protocol High transfection efficacy in the presence of serum Expand your RNAi application with a reagent optimized for delivery of both siRNA and plasmid Reproducible transfection results Works well for standard reverse transfection and high-throughput applications 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 MDA-MB Cells (Breast Adenocarcinoma Cells, HTB-26) Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in the MDA-MB-453 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).

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for MDA-MB Cells (Breast Adenocarcinoma Cells, HTB-26) Data Figure 2. Protein expression of Cyclophilin B in MD-MB cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into MDA-MB 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.

MDA-MB Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for MDA-MB Cells (Breast Adenocarcinoma Cells, HTB-26) MDA-MB Transfection Kit Benefits Pre-optimized transfection protocol for MDA-MB 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 MDA-MB 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 MDA-MB 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