Download PPT - Altogen Biosystems

Survey
yes no Was this document useful for you?
   Thank you for your participation!

* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project

Document related concepts

Cre-Lox recombination wikipedia , lookup

Molecular cloning wikipedia , lookup

Artificial gene synthesis wikipedia , lookup

Secreted frizzled-related protein 1 wikipedia , lookup

DNA vaccination wikipedia , lookup

Cell culture wikipedia , lookup

Transformation (genetics) wikipedia , lookup

List of types of proteins wikipedia , lookup

Channelrhodopsin wikipedia , lookup

Cell-penetrating peptide wikipedia , lookup

Vectors in gene therapy wikipedia , lookup

Transcript
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)
Altogen Biosystems offers the Transfection Reagent for PANC-1
Cells
Transfection Reagent among a host of 100+ cell line
specific In Vitro Transfection Kits.
The PANC-1 Transfection Reagent is a proprietary cationic lipids
formulation that enhances lipid mediated transfection, and it has
been developed to provide high transfection efficiency with PANC1 cells.
This cell line has applications for research related to pancreatic
cancer. When cultured in vitro, PANC-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 PANC-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
Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)
The PANC-1 cell line is an abbreviation for “pancreas,”
and originates from pancreatic duct tissue of a 56-year old
Caucasian male patient suffering from epithelioid
carcinoma. The PANC-1 cell line has a population doubling
time of 52 hours and a chromosomal modal number of 63.
The cell line can be used as an in vitro model in
tumorigenicity studies related to non-endocrine pancreatic
cancer. L-asparagine inhibits growth of PANC-1 cells.
FGF2 stimulates the cell line to differentiate into islet like
clusters that produce hormones. The PANC-1 cell line is
negative for estrogen receptor. The cell line has particular
applications for research related to pancreas cancer, but
also serves an excellent transfection host.
High Density PANC-1 Cells
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 > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)
PANC-1 Transfection Protocol
1. Plate 10,000 - 15,000 PANC-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 PANC-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 PANC-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
Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)
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 > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)
PANC-1 Transfection Kit Product Details
•
Proprietary 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 > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)
Data
Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in PANC-1 cells transfected by
Cyclophilin B siRNA or non-silencing siRNA control following the recommended transfection protocol.
Cyclophilin mRNA expression levels were measured 48 hours post-transfection. 18S rRNA levels were
used to normalize the Cyclophilin B data. Values are normalized to untreated sample. Data are
presented as means ± SD (n=3).
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)
Data
Figure 2. Protein expression of Cyclophilin B in PANC-1 cells. DNA plasmid expressing Cyclophilin B or siRNA
targeting Cyclophilin B were transfected into PANC-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
Products > PANC-1 Transfection Reagent (Non-endocrine Pancreatic Cancer)
PANC-1 Transfection Kit Benefits
• Pre-optimized transfection protocol for PANC-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 PANC-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 PANC-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]