Transcript PPT - Altogen Biosystems

Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
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DLD-1 Transfection Reagent (Colon Cancer, CCL-221)
Altogen Biosystems offers the DLD-1 Transfection Reagent
among a host of 100+ cell line specific In Vitro Transfection Kits.
DLD-1 Transfection Reagent is an advanced formulation of lipid
based reagent, and it has been developed to provide high
transfection efficiency with DLD-1 and other colon cancer cell
lines.
This cell line is a good host for studying colon cancer and related
areas of study. When cultured in vitro, DLD-1 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 DLD-1 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
DLD-1 Transfection Reagent (Colon Cancer, CCL-221)
DLD-1 Cell Line Description and Applications
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DLD-1 Cells
molecular-cancer.biomedcentral.com
The DLD-1 cell line originates from the epithelial
tumorigenic tissue of a Duke type C tumor from
an adult male patient. The patient suffered from
colorectal adenocarcinoma. In regards to
oncogenes, the DLD-1 cell line is negative for
ros, abl, and src, but positive for myb, ras, fos,
sis and p53. DLD-1 is similar to the HCT-15 cell
line as both cells originate from the same slone.
The DLD-1 cell line has applications for
research related to colon 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
DLD-1 Transfection Reagent (Colon Cancer, CCL-221)
DLD-1 Transfection Protocol
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1. Plate 10,000 - 15,000 DLD-1 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 DLD-1 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 DLD-1 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
DLD-1 Transfection Reagent (Colon Cancer, CCL-221)
General Lipoplex-mediated Transfection Mechanism of Action
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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
DLD-1 Transfection Reagent (Colon Cancer, CCL-221)
DLD-1 Transfection Kit Product Details
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•
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 >
DLD-1 Transfection Reagent (Colon Cancer, CCL-221)
Data
Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the DLD-1 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 >
DLD-1 Transfection Reagent (Colon Cancer, CCL-221)
Data
Figure 2. Protein expression of GAPDH in DLD-1 cells. DNA plasmid expressing GAPDH or siRNA targeting
GAPDH were transfected into DLD-1 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
DLD-1 Transfection Reagent (Colon Cancer, CCL-221)
DLD-1 Transfection Kit Benefits
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• Pre-optimized transfection protocol for DLD-1 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 DLD-1 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 DLD-1 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]