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

Gene expression wikipedia , lookup

Deoxyribozyme wikipedia , lookup

Secreted frizzled-related protein 1 wikipedia , lookup

Cre-Lox recombination wikipedia , lookup

Artificial gene synthesis wikipedia , lookup

Molecular cloning 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 > AGS Transfection Reagent (Gastric Adenocarcinoma, CRL1739)
Altogen Biosystems offers the AGS Transfection Reagent among
a host of 100+ cell line specific In Vitro Transfection Kits.
AGS Transfection Reagent is an advanced formulation of a
polymer based reagent, and it is developed to provide high
transfection efficiency with AGS and other gastric adenocarcinoma
cell lines.
This cell line is a good host for studying stomach cancer and
human infections related to the stomach.
Other applications include drug discovery, gene expression
studies, molecular and cell biology research applications.
Purchase AGS 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 > AGS Transfection Reagent (Gastric Adenocarcinoma, CRL1739)
AGS Cell Line Description and Applications
The AGS cell line is derived from the stomach
tissue of a Caucasian female who had
adenocarcinoma or stomach cancer, aged 54
years from 1979. No therapy was administered to
the patient before the cells were taken. The AGS
cell line is tumorigenic and shows epithelial cellular
morphology and this line has the potential to be
used in research related to different types of
stomach cancer.
AGS Cell Line
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 > AGS Transfection Reagent (Gastric Adenocarcinoma, CRL1739)
AGS Transfection Protocol
1. Plate 10,000 - 15,000 AGS 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 AGS 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 AGS 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 > AGS Transfection Reagent (Gastric Adenocarcinoma, CRL1739)
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 > AGS Transfection Reagent (Gastric Adenocarcinoma, CRL1739)
AGS 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 > AGS Transfection Reagent (Gastric Adenocarcinoma, CRL1739)
Data
Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the AGS 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 > AGS Transfection Reagent (Gastric Adenocarcinoma, CRL1739)
Data
Figure 2. Protein expression of GAPDH in AGS cells. DNA plasmid expressing GAPDH or siRNA targeting
GAPDH were transfected into AGS 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 > AGS Transfection Reagent (Gastric Adenocarcinoma, CRL1739)
AGS Transfection Kit Benefits
• Pre-optimized transfection protocol for AGS 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 AGS 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 AGS 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]