abm | AT1A Knockout Immortalized Mouse Renal Proximal Tubule Cell Line | T0626

abm | AT1A Knockout Immortalized Mouse Renal Proximal Tubule Cell Line | T0626

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 AT_1A and AT_1B. The AT1A 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

Biosafety:

II

Organism:

C57B/6 mouse with AT_1A^-/- genotype

Source Organ:

Kidney

Growth Properties:

Adherent

Morphology:

Cobblestone

Clones:

N/A

Passage Number:

N/A

Population Doner:

N/A

Seeding Density:

Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.

Markers:

N/A

Applications:

For Research Use Only

Doner Gender:

N/A

Donor Ethnicity:

N/A

Knockdown Method:

N/A

Induction:

N/A

Overexpression:

N/A

Freeze Thaw:

N/A

Propagation:

The base medium for this cell line is Prigrow IV medium available at abm

Preservation:

1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO. 2. Storage Temperature: Liquid nitrogen vapour phase.

Quality Control:

(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.

Tumorgenicn:

N/A

Shipping Conditions:

Dry Ice

Storage Contidions:

-180°C