Sensitive bioassay for the detection and quantification of angiotensin II in tissue culture medium.
We have developed a sensitive, high-throughput bioassay to quantify angiotensin II in culture medium. Using Chinese hamster ovary cells that stably express a transfected angiotensin II receptor as target cells, we demonstrated that an agonist-stimulated myelin basic protein kinase response can be used as a basis of quantitative bioassay for angiotensin II. The assay permits detection of as little as 10 pg of angiotensin II in 1 mL of medium and offers an excellent alternative to HPLC and radioimmunoassay. This approach may also be applicable for quantification of other peptide hormones or growth factors produced by cell cultures.
Lin
C
C
Weis Center for Research, Danville, PA, USA.
Baker
K M
KM
Thekkumkara
T J
TJ
Dostal
D E
DE
D000804:Angiotensin II / Q000032:analysis*; D057911:Angiotensin Receptor Antagonists; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D001713:Biphenyl Compounds; D016466:CHO Cells / Q000201:enzymology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D006224:Cricetinae; D003470:Culture Media; D016895:Culture Media, Serum-Free; D038362:Glycogen Synthase Kinase 3; D007093:Imidazoles; D019808:Losartan; D051381:Rats; D011945:Receptors, Angiotensin / Q000235:genetics; D012680:Sensitivity and Specificity; D013777:Tetrazoles; D014162:Transfection
D057911:Angiotensin Receptor Antagonists; D001713:Biphenyl Compounds; D003470:Culture Media; D016895:Culture Media, Serum-Free; D007093:Imidazoles; D011945:Receptors, Angiotensin; D013777:Tetrazoles; D000804:Angiotensin II; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D038362:Glycogen Synthase Kinase 3; D019808:Losartan