AT1A/AT1B Double Knockout Immortalized Mouse Renal Proximal Tubule Cell Line
|T0628||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. Even so, there is an incomplete understanding of the AT1 receptors AT1A and AT1B. The AT1B 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), AT1B -/- (Cat. No. T0627) and AT2 -/- (Cat. No. T0625), 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 confirm AT receptor genotype. (2) Electrical conductivity was measured to compare immortalized cells relative to non-immortalized counterparts. (3) Immunostaining was performed to determine localization of proteins associated with epithelial cells and assess morphology. (4) Changes in short circuit current were quantified to compare immortalized cell cotransporters to non-immortalized ones. (5) Western blotting was performed to detect the presence of AT receptor proteins. (6) Live cell microscopy of AngII-mediated β-arrestin 2 translocation was performed to measure 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).
5. 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 application(s). Please contact a technical service representative for more information.
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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."
|Depositor||Case Western Reserve University|
- Li, XC et al. "Mechanisms of AT1a receptor-mediated uptake of angiotensin II by proximal tubule cells: a novel role of the multiligand endocytic receptor megalin" Am J Physiol Renal Physiol. 307(2):F222-33 (2014). PubMed: 24740791.
- Li, XC et al. "Role of caveolin 1 in AT1a receptor-mediated uptake of angiotensin II in the proximal tubule of the kidney" Am J Physiol Renal Physiol. 307(8):F949-61 (2014). PubMed: 25164083 .
- Li, XC et al. "Novel signaling mechanisms of intracellular angiotensin II-induced NHE3 expression and activation in mouse proximal tubule cells" Am J Physiol Renal Physiol. 303(12):F1617-28 (2012). PubMed: 23034941 .
- Li, XC et al. "AT1 receptor-mediated uptake of angiotensin II and NHE-3 expression in proximal tubule cells through a microtubule-dependent endocytic pathway" Am J Physiol Renal Physiol. 297(5):F1342–F1352 (2009). PubMed: PMC2781332.
- Woost, PG et al. "Strategy for the development of a matched set of transport-competent, angiotensin receptor-deficient proximal tubule cell lines." In Vitro Cell Dev Biol Anim 42(7):189-200 (2006). PubMed: 16948500 .