Transcript PPT - Altogen Biosystems

Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > PC-3
Transfection Reagent (Prostate Cancer Cells)
Altogen Biosystems offers the PC-3 Transfection Reagent among
a host of 100+ cell line specific In Vitro Transfection Kits.
The PC-3 Transfection Reagent is an advanced formulation of a
lipid based reagent, and it has been developed to provide high
transfection efficiency with the PC-3 cell line.
This cell line is considered to be a classic cell line for studying
prostate cancer. When cultured in vitro, PC-3 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 PC-3 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 > PC-3
Transfection Reagent (Prostate Cancer Cells)
PC-3 Cell Line Description and Applications
High Density PC-3 cells
Atcc.org
The PC-3 cell line originates from the bone marrow
metastasis of a 62-year-old Caucasian man with
grade IV prostate cancer post mortem. The patient
was treated with androgen suppression therapy.
This cell line has a high metastatic potential and
research indicates that the cells will form tumors in
nude mice. The PC-3 cell line expresses Prostate
Specific Antigen and has a low amount of
testosterone-5-alpha reductase activity. This cell
line has unique applications for research related to
cancer and molecular and cellular biology due to its
high metastatic potential. The PC-3 cell line is also
considered to be a classic cell line in the study of
prostate cancer.
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 > PC-3
Transfection Reagent (Prostate Cancer Cells)
PC-3 Transfection Protocol
1. Plate 10,000 - 15,000 PC-3 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 PC-3 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 PC-3 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 > PC-3
Transfection Reagent (Prostate 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 > PC-3
Transfection Reagent (Prostate Cancer Cells)
PC-3 Transfection Kit Product Details
•
Cationic lipids formulation
•
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  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > PC-3
Transfection Reagent (Prostate Cancer Cells)
Data
Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the PC3 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 > PC-3
Transfection Reagent (Prostate Cancer Cells)
Data
Figure 2. Protein expression of GAPDH in PC-3 cells. DNA plasmid expressing GAPDH or siRNA targeting
GAPDH were transfected into PC-3 cells following Altogen Biosystems transfection protocol. At 72 hours posttransfection 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 > PC-3
Transfection Reagent (Prostate Cancer Cells)
PC-3 Transfection Kit Benefits
• Pre-optimized transfection protocol for PC-3 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 PC-3 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 PC-3 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]