Isolation and Bioassay of AT1-AA

The female hAogen x male hRen (MDC, Berlin) transgenic rats (MDC, Berlin) were used as the source of rat AT1-AA. This model develops hypertension associated with the AT1-AA. On day 18 of gestation blood was collected and immunoglobulin was isolated from one ml of serum by specific anti-ratIgG column purification. AT1-AA was purified from rat IgG by epitope binding to the amino acid sequence corresponding to the second extracellular loop of the AT1 receptor covalently linked to Sepharose 4B CNBr-activated gel. Unbound IgG was washed away and bound IgG was eluted with 3 M potassium thiocyanate. AT1-AA activity was measured utilizing a bioassay that evaluates the beats/minute (bpm) of neonatal cardiomyocytes in culture3 (link),12 (link).

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LaMarca B., Parrish M., Ray L.F., Murphy S.R., Roberts L., Glover P., Wallukat G., Wenzel K., Cockrell K., Martin JN J.r., Ryan M.J, & Dechend R. (2009). Hypertension in response to autoantibodies to the angiotensin II type I receptor (AT1-AA) in pregnant rats: Role of endothelin-1. Hypertension, 54(4), 905-909.

Publication 2009

Amino acid sequence Bioassay Blood Cardiomyocytes Cnbr Epitope Female Gestation Hypertension Immunoglobulin Male Neonatal Potassium thiocyanate Sepharose 4b Serum Transgenic rats

Corresponding Organization :

Other organizations : University of Mississippi Medical Center, Charité - Universitätsmedizin Berlin

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Variable analysis

independent variables

The female hAogen x male hRen (MDC, Berlin) transgenic rats (MDC, Berlin)

dependent variables

AT1-AA activity measured utilizing a bioassay that evaluates the beats/minute (bpm) of neonatal cardiomyocytes in culture

control variables

Day 18 of gestation

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