A30P aSYN-IRES-GFP Stable LUHMES Cell Line
Cat. No.
T6454
Unit
1x10⁶ cells / 1 ml
Price
Inquiry
Worried about losing your cells due to growth or thawing difficulties, or even a random freezer breakdown? Enjoy peace of mind knowing that you can be covered under abm's Cell Line Insurance.
Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing institution.
For for-profit organizations, please contact quotes@abmgood.com for pricing.
Specifications
| Description |
This cell line, the A30P aSYN-IRES-GFP Stable LUHMES Cell Line, was generated through lentivral infection. These cells stably express A30P aSyn and are suitable for investigations focusing on neurotoxicity and the identification of targets for the treatment/prevention of neurodegenerative disorders. abm also offers: A53T aSYN-IRES-GFP Stable LUHMES Cell Line (T6455), Wild Type aSYN-IRES-GFP Stable LUHMES Cell Line (T6456) IRES-GFP Stable LUHMES Cell Line (T6457) |
| Cat. No. | T6454 |
| Name | A30P aSYN-IRES-GFP Stable LUHMES Cell Line |
| Unit | 1x10⁶ cells / 1 ml |
| Price | Inquiry |
| Category | Stable Cell Lines |
| Cell Type | Stable Cell Lines |
| Organism | Human (H. sapiens) |
| Tissue | Brain |
| Donor History | 8-week-old fetal human ventral mesencephalon |
| Cell Morphology | Epithelial |
| Growth Properties | Adherent, epithelial |
| Expression Profile |
A30P aSyn, GFP, Tetracycline resistance |
| Expression | A30P aSyn, GFP, Tetracycline resistance |
| Propagation Method |
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 or 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). For information regarding cell differentiation, please refer to the Differentiation Protocol PDF under the Documents Tab. |
| Growth Conditions | 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 or 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. Change media every 2-3 days. Do not let media colour change to orange-yellow. The cells will form round clumps instead of a monolayer when stressed. It is recommended to subculture when cells are grown to a cell density of 50-60%. Carbon dioxide (CO2): 5%, Temperature: 37.0°C. 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). For information regarding cell differentiation, please refer to the Differentiation Protocol PDF under the Documents Tab. |
| Subculture Protocol |
Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent. 1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions. |
| QC |
1) Western blot; 2) Immunocytochemistry |
| Application | Research Use Only. |
| Storage Condition | Vapor phase of liquid nitrogen, or below -130°C. |
| BioSafety | II |
| Disclaimer |
1. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each cell line. If you have any questions regarding this, please contact us at licensing@abmgood.com. 2. All test parameters provided in the CoA are conducted using abm's standardized culture system and 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. 3. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of order placement. 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. 4. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic or any other non-RUO application(s). 5. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature are provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 6. 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". |
| Depositor | Georg-August-Universität Göttingen |
STR
There are no STR for this product yet!
FAQs
| I want to make sure these cells express my gene of interest before I decide to buy the cell line. Can you provide a sample so this can be tested? | |
|
We do not perform extensive downstream characterization, or gene expression profiling of our cell lines. However, you can check the ‘References’ tab to see if a publication is available, or you can contact us to see if RNA or cell lysates are available for your cell line of interest. Please directly contact us for further information at quotes@abmgood.com.
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| Does abm's PriGrow series medium contain antibiotics? | |
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No, the medium does not contain antibiotics and needs to be supplemented by the end-user as desired.
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| Do I need to use abm's media and Applied Cell Extracellular Matrix (G422) to culture my cells? | |
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We strongly encourage using the recommended media and culture conditions described in the "Growth Conditions" section as they have been optimized for cell growth. Other supplier's media and coating conditions have not been tested.
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| What is the difference between Applied Cell Extracellular Matrix (G422) and Collagen Coating? | |
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The main component of our Applied Cell Extracellular Matrix (G422) is type I collagen specifically. For more information on abm's Applied Cell Extracellular Matrix please visit: http://www.abmgood.com/Applied-Cell-Extracellular-Matrix-G422.html.
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| Do I have to use T25 ECM-coated flasks for growing the cells? | |
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We strongly recommend thawing cells in the T25 flasks specified on the cell-specific product page to ensure optimal post-thaw recovery of the frozen cells before testing other culture vessels.
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| If I want to plate these cells to multi-well plates (e.g. 96 well plates) or dishes, how should I prepare the plates? | |
| What percentage of Trypsin-EDTA should be used to subculture cells? Can I use Trypsin-EDTA containing phenol red? | |
|
The standard concentration used for subculturing is 0.25% Trypsin-EDTA, available at abm (Cat. No. TM050). Yes, Trypsin-EDTA containing phenol red can be used as it does not interfere with trypsinization.
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| How often do I need to change the media? | |
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Media should be changed every 2-3 days or as specified within the recommended growth conditions.
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| Why are these cells classified as biosafety level II? | |
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We follow the CDC-NIH recommendations that all mammalian sourced products should be handled at the Biological Safety Level 2 to minimize exposure of potentially infectious products. This information can be found in 'Biosafety in Microbiological and Biomedical Laboratories' (1999). Your institution's Safety Officer or Technical Services will be able to make the call as to whether BioSafety Level I is possible with these cells at your site, if desired.
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| How long can I store frozen vials? | |
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Cells that are properly frozen using an effective cryoprotective agent can be stored in liquid nitrogen vapor phase (or below -130°C) indefinitely without affecting viability.
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| What is the recommended storage temperature for cells? | |
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In general, if you received:
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| Can I substitute heat-inactivated FBS with FBS or vice versa? | |
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FBS and heat-inactivated FBS are different in their composition; they cannot be substituted for one another.
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| My cells are not detaching, what method do you recommend to trypsinize the cells? | |
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| How are live cells shipped? | |
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Live cells are shipped as a live culture in a T12.5 flask. Cells are seeded at an optimal confluency to avoid metabolic waste build up during transport. In addition, we thoroughly test our cell lines for functional freeze-thaw recovery. For more information on how to handle live cells, please view our detailed instructions here (https://www.abmgood.com/uploads/document/Cell_Handling_Instructions_Upon_Arrival_150623.pdf).
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| Why are cells not attaching and forming clumps after subculturing with trypsin? | |
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Cells form clumps and display attachment issues when the trypsinization agent is not effectively neutralized. To completely neutralize the trypsin agent, add an equal amount of complete growth media prior to centrifugation. Subtle changes in culture conditions, particularly in pH and the quality of serum used in the growth medium, may also affect the amount of clumping exhibited by the cells.
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| Why are my cells forming clumps after plating? | |
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It is important to ensure cell clumps are broken up through gentle re-suspension by pipetting up and down several times after pelleting cells by centrifugation. This will ensure a homogenous single cell suspension for even plating.
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| Do I need to add the selection drug to the complete growth medium? | |
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We recommend maintaining selection pressure using the drug concentration specified in the Growth Conditions section.
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| Where can I find the CoA for my product? | |
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Certificates of analysis can be found here (https://www.abmgood.com/CoA-Library.html).
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References
There are no references for this product yet!
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