A30P aSYN-IRES-GFP Stable LUHMES Cell Line
|T6454||1x106 cells / 1.0 ml|
Through lentiviral infection, the A30P aSYN-IRES-GFP Stable LUHMES Cell Line was generated. The cells stably express A30P aSyn. This cell line can be used to study neurotoxicity and to identify targets for treatment/prevention of neurodegenerative disorders.
|Species||Human (H. sapiens)|
For Research Use Only
|Unit quantity||1x106 cells / 1.0 ml|
|Cell Type||Drug Discovery Cell Lines|
A30P aSyn, GFP, Tetracycline resistance
Grow the cells in culture vessel pre-coated with 50 μg/ml poly-L-ornithine (PLO) (TM062) and 1 μg/ml fibronectin (EMD Millipore; Cat. FC010) in H2O for at least 3 hours at 37°C. Do not grow these cells in culture vessels with surface areas equal to less than 12.5 cm2. These cells do not grow in 6-well, 24-well, 48-well, or 96-well plates.
The base medium for this cell line is Advanced DMEM/F12 (Gibco;12634010). To make the complete growth medium, add the following components to the base medium: N2 supplement (ThermoFisher Scientific) to a final concentration of 1X, L-glutamine (G275) to a final concentration of 2 mM, and Recombinant Human FGF2 (Z101455) to a final concentration of 40 ng/ml. Cells may be grown in the presence of 1 μg/ml Tetracycline.
To subculture the cells, use TrypLE Express. After adding TrypLE Express, incubate the cells at 37.0°C incubator for 2-3 minutes and agitate the culture vessel until 90% of the cells have detached. Immediately neutralize the trypsin using Trypsin Neutralizing Solution (Lonza, Cat. CC-5002). Add Trypsin Neutralizing Solution the same volume of trypsin used. Centrifuge at 200x g for 2-3 minutes. Aspirate supernatant and resuspend cells in complete media. Plate cells into pre-warmed PLO-fibronectin-coated vessels at 37.0°C. Avoid subculturing if the cells appear stressed. Note: If leaving the cells over the weekend (or more than 2 days), make sure to do a high split ratio (1:4 to 1:5).
To differentiate the cells into neurons, change the medium to differentiation medium after the cells have grown to a density of 40-50%.
1) Western blot; 2) Immunocytochemistry
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2. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected].
3. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.
4. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).
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7. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
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