Characterization of a panel of six beta2-adrenergic receptor antibodies by indirect immunofluorescence microscopy.

Respiratory Research
Yulia A KoryakinaRichard C Kurten

Abstract

The beta2-adrenergic receptor (beta2AR) is a primary target for medications used to treat asthma. Due to the low abundance of beta2AR, very few studies have reported its localization in tissues. However, the intracellular location of beta2AR in lung tissue, especially in airway smooth muscle cells, is very likely to have a significant impact on how the airways respond to beta-agonist medications. Thus, a method for visualizing beta2AR in tissues would be of utility. The purpose of this study was to develop an immunofluorescent labeling technique for localizing native and recombinant beta2AR in primary cell cultures. A panel of six different antibodies were evaluated in indirect immunofluorescence assays for their ability to recognize human and rat beta2AR expressed in HEK 293 cells. Antibodies capable of recognizing rat beta2AR were identified and used to localize native beta2AR in primary cultures of rat airway smooth muscle and epithelial cells. beta2AR expression was confirmed by performing ligand binding assays using the beta-adrenergic antagonist [3H] dihydroalprenolol ([3H]DHA). Among the six antibodies tested, we identified three of interest. An antibody developed against the C-terminal 15 amino acids of the human beta2A...Continue Reading

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Citations

May 24, 2019·Pain Medicine : the Official Journal of the American Academy of Pain Medicine·Peter D DrummondPhilip M Finch
Dec 19, 2020·Molecular Therapy. Methods & Clinical Development·Alexander KondrashovChris Denning

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Methods Mentioned

BETA
confocal microscopy
transfection

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