KCC2 Stable HEK293 Cell Line
|T3038||1x106 cells / 1.0 ml|
Potassium-chloride transporter member 5 (SLC12A5, also known as KCC2) is a neuron-specific K-Cl symporter responsible for maintaining ion homeostasis in the central nervous system. The electrochemical chloride gradient established by KCC2 activity is critical in regulating postsynaptic inhibition, as well as in protecting cells from stimulation-induced excitotoxicity. The KCC2 Stable HEK293 cell line stably over-expresses the neuronal-specific K-Cl co-transporter isoform KCC2. In contrast to KCC1, where its activity is stimulated by osmotic swelling, KCC2 activity is increased when stimulated with NEM (N-ethylmaleimide). This cell line is suitable for central nervous system studies in the potential roles of KCC2 in ion homeostatsis.
|Species||Human (H. sapiens)|
|Species description||Homo sapiens|
|Seeding Density||30,000 - 50,000 cells/cm2, Split ratio: 1:2 or 1:3|
|Population Doubling Time||30 - 40 hours|
For Research Use Only
|Unit quantity||1x106 cells / 1.0 ml|
|Caution||For Research Use Only|
|Cell Type||Drug Discovery Cells|
Use of PriCoatTM T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.
The base medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10% and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. To culture the cells in the presence of selection, add 2 µg/ml Puromycin (G264) to the complete media.
|Preservation Protocol||1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.|
2. Storage Temperature: Liquid nitrogen vapour phase.
1) Cell line was screened using the 86Rb influx assay to determine KCC2 expressed at cell surface; 2) Western blot was used to examine protein expression
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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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- Delpire, E et al. "Small-molecule screen identifies inhibitors of the neuronal K-Cl cotransporter KCC2" Proc Natl Acad Sci USA 106(13):5383-5388 (2009). DOI: 10.1073/pnas.0812756106. Epub 2009 Mar 11.
- Delpire, E et al. "Further optimization of the K-Cl cotransporter KCC2 antagonist ML077: development of a highly selective and more potent in vitro probe" Bioorg Med Chem Lett 22(14):4532-4535 (2012). DOI: 10.1016/j.bmcl.2012.05.126. Epub 2012 Jun 7.
- Medina, I et al. "Current view on the functional regulation of the neuronal K(+)-Cl(-) cotransporter KCC2" Front Cell Neurosci 8:27 (2014). DOI: 10.3389/fncel.2014.00027. eCollection 2014.