Transcript PPT - Altogen Biosystems

Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > DU145
Transfection Reagent (Prostate Carcinoma Cells)
Altogen Biosystems offers the DU145 Transfection Reagent
among a host of 100+ cell line specific In Vitro Transfection
Kits.
The DU145 Transfection Reagent is an advanced lipid
formulation reagent, and it has been developed to provide
high transfection efficiency with the DU145 cell line.
This cell line is a good host for studying prostate cancers
and cell and molecular biology. When cultured in vitro,
DU145 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 DU145 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 > DU145
Transfection Reagent (Prostate Carcinoma Cells)
DU145 Cell Line Description and Applications
High Density DU145 cells
The DU145 cell line originates from human prostate cancer
cells is considered to be a “classical” prostate cancer cell
line. The cell line displays epithelial cell morphology and
grows as a monolayer. The DU145 cell line has a moderate
potential for metastasis, is not sensitive to hormones and
does not express prostate specific antigen (PSA). The cell
line is mildly positive for acid phosphatase and analysis of
the DU145 cell line shows tonofilaments, desmosomes and
microvilli. Research indicates that glucosamine suppresses
DU145 growth. The DU145 and PC3 cell lines are used to
study prostate cancer, a prevalent cancer, particularly
among men over the age of 50.
atcc.org
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 > DU145
Transfection Reagent (Prostate Carcinoma Cells)
DU145 Transfection Protocol
1. Plate 10,000 - 15,000 DU145 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 DU145 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 DU145 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 > DU145
Transfection Reagent (Prostate Carcinoma 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 > DU145
Transfection Reagent (Prostate Carcinoma Cells)
DU145 Transfection Kit Product Details
•A cationic lipid formulation
•Transfection protocols provided for transfection of proteins, DNA, mRNA, siRNA, shRNA and
microRNA
•Transfection Enhancer reagent provided with the kit
•Produce higher level of recombinant protein expression with minimal disruption of normal cell function
•Generate physiologically relevant data you can trust
•Effective for plasmid DNA/siRNA co-transfection
•Easy-to-use transfection protocol with reproducible results
•Low cytotoxicity
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 > DU145
Transfection Reagent (Prostate Carcinoma Cells)
Data
Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into
DU145 cells following the recommended protocol. At 48 hours post-transfection the cells were analyzed
by qRT-PCR for Lamin A/C 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=4).
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > DU145
Transfection Reagent (Prostate Carcinoma Cells)
Data
Figure 2. Protein expression of Lamin A/C in DU145 cells. DNA plasmid expressing Lamin A/C or siRNA
targeting Lamin A/C were transfected into DU145 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 > DU145
Transfection Reagent (Prostate Carcinoma Cells)
DU145 Transfection Kit Benefits
• Pre-optimized transfection protocol for DU145 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 DU145 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 DU145 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]