Immunofluorescence analysis of Adenosine Receptor A1 was performed using 90% confluent log phase HeLa 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 Adenosine Receptor A1 Rabbit Polyclonal Antibody (Product # PA3-041) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization in the membrane. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptides corresponding to the C-terminal portion of human and mouse Adenosine Receptor A1.|
|Storage buffer||whole serum|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500|
* 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.
PA3-041 detects Adenosine Receptor A1 from human and mouse samples. Reactivity is stronger in mouse, and is only recommended for use in overexpressed or recombinant human samples.
PA3-041 has been successfully used in Western blot applications. By Western blot PA3-041 detects a ~34-37 kDa band representing Adenosine Receptor A1 from mouse brain.
The PA3-041 immunogen is a mix of synthetic peptides corresponding to the C-terminal portion of human and mouse Adenosine Receptor A1 residues. The immunizing sequences are: C(309) Q P A P P I D E D L P E E R P D D(326) for human, and C(309) Q P K P P I E E D I P E E K A D D(326) for mouse.
Adenosine receptors (ARs) are members of the 7-transmembrane domain G-protein-coupled receptor superfamily. Structural, biochemical and pharmacological analyses of the AR genes and protein has led to the discovery of four distinct AR subtypes (A1, A2a, A2b, A3). Activation of ARs mediates several receptor subtype-specific physiological processes that include cardiac rate, smooth muscle tone, platelet aggregation, inflammation, cell growth and death, and neurotransmission.
The A1AR is a glycoprotein that can activate Gi and Go proteins in vitro. In intact cells, agonist occupation of the A1AR has been shown to cause pertussis toxin-sensitive inhibition of adenylyl cyclase activity and, in some systems, a stimulation of phospholipase C resulting in mobilization of intracellular calcium stores. Activation of potassium channels by A1AR has been intensively studied in relation to its dramatic effects on the cardiovascular system. A1AR protein is highly expressed in brain (especially cerebellum, hippocampus, thalamus, and cortex) and spinal cord and in part, modulates neurotransmitter release. In white adipocytes A1AR inhibits lipolysis and stimulates glucose uptake. Other tissues also express A1AR including kidney and testis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.