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

Transformation (genetics) wikipedia , lookup

Cell culture wikipedia , lookup

List of types of proteins wikipedia , lookup

Cell-penetrating peptide wikipedia , lookup

Channelrhodopsin 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 > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163)
Altogen Biosystems offers the BALB/3T3 Transfection Reagent
among a host of 100+ cell line specific In Vitro Transfection Kits.
The BALB/3T3 Transfection Reagent is a nanoparticle-based
liposome formulation reagent that enhances lipid mediated
transfection, and it has been developed to provide high
transfection efficiency with BALB/3T3 cells.
This cell line is a good host for transfection and the assessment of
cellular functionality. When cultured in vitro, BALB/3T3 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 BALB/3T3 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 > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163)
BALB/3T3 Cell Line Description and Applications
The BALB cell line is also known as the 3T3 cell line
(clone A31). The cell line originates from Mus
musculus embryos and was established in 1968. For
optimal BALB cell growth, care must be given to
environment as the BALB cell line is extremely
sensitive to environmental factors. The BALB cell line
is particularly useful for studying mammal embryonic
stem cells such as stem cells. Furthermore, this
BALB/3T3 cell line can serve as a transfection host in
a spectrum of biology research.
High Density BALB/3T3 Cells
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 > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163)
BALB/3T3Transfection Protocol
1. Plate 10,000 - 15,000 BALB/3T3 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 BALB/3T3cells (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 BALB/3T3cells 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 > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163)
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 > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163)
BALB/3T3Transfection Kit Product Details
•
A nanoparticle-based liposome 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 trust4
•
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 > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163)
Data
Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in BALB/3T3cell line 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=6).
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163)
Data
Figure 2. Protein expression of Lamin A/C in BALB/3T3 cells. DNA plasmid expressing Lamin A/C or siRNA
targeting Lamin A/C were transfected into BALB/3T3 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 > Transfection Reagent for BALB/3T3 Cells (Embryo Fibroblast Cells, CCL-163)
BALB/3T3Transfection Kit Benefits
• Pre-optimized transfection protocol for BALB/3T3 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 BALB/3T3 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 BALB/3T3 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]