Label ATTO-488. Maximum absorption 501 nm; maximum fluorescence 523 nm. The fluorescence is excited most efficiently in the 480 – 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 50 µl double distilled water (DDW).
Antibody concentration after reconstitution 1 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 × g 5 min).
Standard quality control of each lot Western blot analysis (unlabeled antibody, #AAR-018), and live cell imaging (labeled antibody).
Applications: FC, ICC, IF, LCI
Direct flow cytometry
Rat GH3 pituitary adenoma cells (10 µg/5x105 cells).
Cell surface detection of α1B-Adrenoceptor in live rat GH3 pituitary adenoma cells:
Cell surface detection of α1B-Adrenoceptor in GH3 cells with Anti-α1B-Adrenergic Receptor (extracellular)-ATTO Fluor-488 Antibody (#AAR-018-AG) (green). Nuclear fluorescence staining of cells using the membrane-permeable DNA dye Hoechst 33342 (blue).
Scientific background
Adrenergic receptors (also called adrenoceptors) are the receptors for the catecholamines adrenaline and noradrenaline (called epinephrine and norepinephrine in the United States). Adrenaline and noradrenaline play important roles in the control of blood pressure, myocardial contractile rate and force, airway reactivity, and a variety of metabolic and central nervous system functions.
Adrenergic receptors are members of the G-protein coupled receptor (GPCR) superfamily of membrane proteins. They share a common structure of seven putative transmembrane domains, an extracellular amino terminus, and a cytoplasmic carboxyl terminus.
Adrenoceptors are divided into three types: α1, α2 and β adrenoceptors. Each type is further divided into at least three subtypes: α1A, α1B, α1D, α2A, α2B, α2C, β1, β2, β3.1,2 They are expressed in nearly all peripheral tissues and in the central nervous system.1,2
All α1-adrenoceptors (α1-ARs) activate phospholipases C and A2.3 In addition to mobilizing intracellular calcium, the α1-ARs have also been shown to activate calcium influx via voltage-dependent and -independent calcium channels.4
α1B-Adrenoceptor populations are greatest in the spleen, kidney, cerebellum, and fetal brain.5
α1B-Adrenoceptor causes contraction of smooth muscle cells and thereby controls vascular tone, blood pressure, and accelerates the development of atherosclerosis.5
Application key:
CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot