AT2 Knockout Immortalized Mouse Renal Proximal Tubule Cell Line
|T0625||1x106 cells / 1.0 ml|
|Description||The proximal tubule is part of the nephron duct system in the kidney, involved in regulating the renin-angiotension system. Cells of this region aid in regulating sodium and volume homeostasis as well as regulating the systemic blood pressure. The protein Angiotension II (Ang II) and its receptors AT1 and AT2 are some of the most studied proteins in this system. The AT2 Knockout Immortalized Mouse Renal Proximal Tubule Cell Line is a conditionally immortalized mouse renal proximal tubule cells isolated from the mouse harboring thermolabile mutation (tsA58) of the simian virus 40 large T antigen. The functional expression of the SV40 large T antigen is induced by culturing the cells in vitro in medium containing IFN γ at a temperature permissive (33°C). At a non-permissive temperature (37°C-39°C), the cells cease to proliferate. When paired up with the wild type (Cat. No. T0624) and other angiotensin receptor (Ang II)-deficient cell lines AT1A -/- (Cat. No. T0626), lines AT1B -/- (Cat. No. T0627) and AT1A -/- AT1B -/- (Cat. No. T0628), these cell lines represent valuable tools to study fluid and electrolyte transportation in the proximal tubule region.|
|Species||Mouse (M. musculus)|
|Species description||Mouse (C57BL/6J)|
|Seeding Density||Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.|
|Applications||For Research Use Only|
|Unit quantity||1x106 cells / 1.0 ml|
|Pharmaceutical Target||GPC Receptors|
|Caution||For Research Use Only|
|Cell Type||Drug Discovery Cells|
Use of PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.The base medium for this cell line is Prigrow IV medium available at abm, Cat. No. TM004. To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 5%, 10nM aldosterone, 50μM L-ascorbic acid 2-phosphate, 4μg/mL dexamethasone, 10ng/mL epidermal growth factor, 5μg/mL insulin, 20nM sodium selenite (Na2SeO3), 5μg/mL transferrin, 1nM L-3,3’,5-triiodothyronine (T3), 10 U/mL recombinant mouse interferon gamma (IFN-?) and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: 33.0°C.
Grow cells on 5% gelatine and 10 mg/mL laminin (diluted 1:1) coated vessels. To differentiate the cells, grow the cells at 37°C-39°C in the absence of IFN-?.
|Preservation Protocol||1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.|
2. Storage Temperature: Liquid nitrogen vapour phase.
|QC||(1) RT-PCR was performed to examine products expressed in mutant AT2 -containing cells. (2) Immunostaining was performed to detect distribution of AT1and AT2, plus assess morphology. (3) Western blotting was performed to confirm presence of AT2. (4)Electrical conductivity and changes in short-circuit currents were measured to determine how similar immortalized cells were to their non-immortalized counterparts. (5)Radioimmunoassays were performed to detect guanylate cyclase levels as a measure of AT2 receptor functionality.|
1. For for-profit organizations and corporations, please contact [email protected] for pricing of this item.
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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|Depositor||Case Western Reserve University|
- Woost, PG et al. "Development of an AT2-deficient proximal tubule cell line for transport studies" In Vitro Cell Dev Biol Anim 43(10):352-60 (2007). PubMed: 17963016 .