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

Molecular cloning 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 > U87
Transfection Reagent (Glioblastoma Cells)
Altogen Biosystems offers the U87 Transfection Reagent among a
host of 100+ cell line specific In Vitro Transfection Kits.
The U87 Transfection Reagent is an advanced formulation of a
lipid based reagent, and it has been developed to provide high
transfection efficiency with the U87 cell line.
This cell line is used for researching cytotoxicity of
chemotherapies as well as for researching gliobastoma. When
cultured in vitro, U87 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 U87 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 > U87
Transfection Reagent (Glioblastoma Cells)
U87 Cell Line Description and Applications
High Density U87 cells
Atcc.org
The U87 cell line is crucial to investigating the toxicity of
chemotherapies aimed at cancer as well as for
investigating the in vitro model of gliobastoma cells.
Stage 4 gliobastoma or gliobastoma multiform (GBM) is a
deadly brain cancer; it is a primary malignant brain tumor
and there are approximately 16,000 new diagnoses
yearly in the United States alone. Furthermore, GBM
only has a 29.6 % one-year survival rate. The U87 cell
line was established from a 44-year old Caucasian
female with Stage 3 GBM. The cell line has epithelial
morphology and has a modal chromosome number of 44.
All of the U87 cells have 12 makers in common and N1,
N6 and N9 were not found.
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 > U87
Transfection Reagent (Glioblastoma Cells)
U87 Transfection Protocol
1. Plate 10,000 - 15,000 U87 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 U87 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 U87 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 > U87
Transfection Reagent (Glioblastoma 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 > U87
Transfection Reagent (Glioblastoma Cells)
U87 Transfection Kit Product Details
•
Two component formulation enhances lipid mediated transfection efficiency
•
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 > U87
Transfection Reagent (Glioblastoma Cells)
Data
Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the U87 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 > U87
Transfection Reagent (Glioblastoma Cells)
Data
Figure 2. Protein expression of GAPDH in U87 cells. DNA plasmid expressing GAPDH or siRNA targeting
GAPDH were transfected into U87 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 > U87
Transfection Reagent (Glioblastoma Cells)
U87 Transfection Kit Benefits
• Pre-optimized transfection protocol for U87 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 U87 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 U87 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]