Transcript PPT - Altogen Biosystems

Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > HMEC Transfection Reagent (Mammary Epithelial, Breast Cancer Cells)
Altogen Biosystems offers the HMEC Cell Transfection Reagent
among a host of 100+ cell line specific In Vitro Transfection Kits.
The HMEC Cell Transfection Reagent is a lipid mediated
formulation, and it has been developed to provide high
transfection efficiency with HMEC.
This cell line is a good host for studying breast cancer and
oncogenesis. When cultured in vitro, HMEC 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 HMEC 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  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > HMEC Transfection Reagent (Mammary Epithelial, Breast Cancer Cells)
HMEC Cell Line Description and Applications
HMEC cells
The HMEC cell line originates from human mammary
epithelial cancer cells. These cells were surgically
removed from glandular tissue from a human female
patient suffering from breast cancer. This cell line
displays cuboidal morphology with a characteristic
pattern of swirls. Research studies regarding breast
tissue indicate that cell cultures derived from breast
cancer tissue are ideal for studying therapeutics or
oncogenesis. The primary application of the HMEC
Transfection Kit is with human mammary epithelial
cancer cells and studying breast cancer. Other
applications of the HMEC cell line include carcinogen
screening and three dimensional culturing.
Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA
Telephone  702 349 6103  Fax  702 989-0841  email  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > HMEC Transfection Reagent (Mammary Epithelial, Breast Cancer Cells)
HMEC Transfection Protocol
1. Plate 10,000 - 15,000 HMEC 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 HMEC 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 HMEC 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  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > HMEC Transfection Reagent (Mammary Epithelial, Breast Cancer 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  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > HMEC Transfection Reagent (Mammary Epithelial, Breast Cancer Cells)
HMEC Transfection Kit Product Details
•
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  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > HMEC Transfection Reagent (Mammary Epithelial, Breast Cancer Cells)
Data
Figure 1. SiRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into
HMEC cells following the recommended protocol. At 48 hours post-transfection the cells were analyzed
by qRT-PCR for gene expression levels. 18S rRNA levels were used to normalize the Lamin A/C data.
Values are normalized to untreated sample. Data are means ± SD (n=3).
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > HMEC Transfection Reagent (Mammary Epithelial, Breast Cancer Cells)
Data
Figure 2. Protein expression of Lamin A/C in HMEC cells. DNA plasmid expressing Lamin A/C or siRNA
targeting Lamin A/C were transfected into HMEC 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.
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > HMEC Transfection Reagent (Mammary Epithelial, Breast Cancer Cells)
HMEC Transfection Kit Benefits
• Pre-optimized transfection protocol for HMEC 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 HMEC 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 HMEC 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  [email protected]