Immunofluorescence analysis of beta-Arrestin 1 was done on 70% confluent log phase T47D cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with beta-Arrestin 1 (ZA005) Mouse Monoclonal Antibody (395000) at 2ug/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Synthetic peptide derived from the C-terminal region of human beta-arrestin 1|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Miscellaneous PubMed (MISC)||See 1 publications below|
Beta Arrestin 1 is a member of a family of proteins widely expressed but especially abundant in the central nervous system. Serving as an adaptor or scaffold maolecule, beta Arrestin 1 is essential for mitogenic signalling and mediates agonist-dependent desensitization and internalization of Gprotein-coupled receptors (GPCRs, e.g., beta 2-adrenergic receptor). After binding to their ligand and interacting with heterotrimeric G proteins, GPCRs are phosporylated by G-protein receptor kinases (GRKs) on serine residues. Beta Arrestin 1 in the cytosol is phosphorylated by ERK1 and 2 on serine412 in a negative feedback mechanism and binds to the phosphorylated receptors at the plasma membrane. Serine 412 is then dephosphorylated and the GPCRs are internalized, leading to activation of the Ras, Raf, ERK1 and 2 signaling pathway.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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