Download PPT - Altogen Biosystems

Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > 786-O Transfection Reagent (Prostate Carcinoma, CRL-1932)
Altogen Biosystems offers the 786-O Transfection Reagent
among a host of 100+ cell line specific In Vitro Transfection Kits.
786-O Transfection Reagent is an advanced formulation of a
biodegradable polymer-based reagent, and it is developed to
provide high transfection efficiency with 786-O and other prostate
carcinoma cell lines.
This cell line is a good host for studying prostate cancer and
human infections related to the prostate. When cultured in vitro,
786-O 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 786-O 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 > 786-O Transfection Reagent (Prostate Carcinoma, CRL-1932)
786-O Cell Line Description and Applications
786-O Cell Line
www.clsgmbh.de
The 786-O cell line originates from primary clear
cell adenocarcinoma kidney tissue from a
Caucasian 58-year old male patient with renal cell
adenocarcinoma. The 786-O cell line produces
parathyroid hormone (PTH) and displays epithelial
cell morphology. The PTH that is produced by the
786-O cell line is identical to the peptides that are
produced by lung and breast tumors. The cell has
applications for several areas of research in
cellular biology and it is an excellent transfection
host.
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 > 786-O Transfection Reagent (Prostate Carcinoma, CRL-1932)
786-O Transfection Protocol
1. Plate 10,000 - 15,000 786-O 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 786-O 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 786-O 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 > 786-O Transfection Reagent (Prostate Carcinoma, CRL-1932)
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 > 786-O Transfection Reagent (Prostate Carcinoma, CRL-1932)
786-O 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 > 786-O Transfection Reagent (Prostate Carcinoma, CRL-1932)
Data
Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the 786-O 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 GAPD 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).
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > 786-O Transfection Reagent (Prostate Carcinoma, CRL-1932)
Data
Figure 2. Protein expression of GAPDH in 786-O cells. DNA plasmid expressing GAPDH or siRNA targeting
GAPDH were transfected into 786-O 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 > 786-O Transfection Reagent (Prostate Carcinoma, CRL-1932)
786-O Transfection Kit Benefits
• Pre-optimized transfection protocol for 786-O 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 786-O 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 786-O 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]