Download PPT - Altogen Biosystems

Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > HCAEC Transfection Reagent (Coronary Artery Endothelial)
Altogen Biosystems offers the HCAEC Cell Transfection Reagent
among a host of 100+ cell line specific In Vitro Transfection Kits.
The HCAEC Cell Transfection Reagent is a nanoparticle-based
liposome mediated formulation, and it has been developed to
provide high transfection efficiency with HCAEC.
This cell line is a good host for studying the circulatory system and
the heart as well as a range of areas within the field of biomedical
therapeutics and biology. When cultured in vitro, HCAEC 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 HCAEC 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 > HCAEC Transfection Reagent (Coronary Artery Endothelial)
HCAEC Cell Line Description and Applications
HCAEC cells
lonza.com
The HCAEC or human coronary artery endothelial cell
line originates from endothelial cells taken from a
human coronary artery. HCAEC cells were isolated
from both the left and right coronaries as well as the
circumflex branches and anterior descending coronary
from a single donor. These cells are critical to coronary
blood flow as well as cardiac functions. The cell line
displays epithelial cell morphology. The HCAEC cell line
can be used to study the circulatory system and the
heart as well as related diseases such as thrombosis,
hypertension and atherosclerosis. The HCAEC cell line
can also be used to study vasodilators.
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 > HCAEC Transfection Reagent (Coronary Artery Endothelial)
HCAEC Transfection Protocol
1. Plate 10,000 - 15,000 HCAEC 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 HCAEC 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 HCAEC 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 > HCAEC Transfection Reagent (Coronary Artery Endothelial)
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 > HCAEC Transfection Reagent (Coronary Artery Endothelial)
HCAEC Transfection Kit Product Details
•
A nanoparticle-based liposome formulation
•
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 > HCAEC Transfection Reagent (Coronary Artery Endothelial)
Data
Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the cells transfected with siRNAs
targeting GAPD 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=3).
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > HCAEC Transfection Reagent (Coronary Artery Endothelial)
Data
Figure 2. Protein expression of GAPDH in HCAEC cells. DNA plasmid expressing GAPDH or siRNA targeting
GAPDH were transfected into HCAEC 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 > HCAEC Transfection Reagent (Coronary Artery Endothelial)
HCAEC Transfection Kit Benefits
• Pre-optimized transfection protocol for HCAEC 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 HCAEC 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 HCAEC 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]