Immunofluorescent analysis of Adenosine Receptor A2a (green) showing staining in the cytoplasm of U251 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an Adenosine Receptor A2a polyclonal antibody (Product # PA1-042) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Dog, Human, Mouse, Rat|
|Published species reactivity||Dog, Rat, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues E(373) S H G D M G L P D V E L L S H E L K(391) of canine A2aAR.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:1,000|
* 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.
|Western Blot (WB)||See 3 publications below|
PA1-042 detects adenosine receptor A2a (A2aAR) in human, rat, mouse and canine tissues. This antibody does not detect other AR subtypes.
PA1-042 has been successfully used in Western blot, ICC/IF, immunohistochemistry and immunoprecipitation procedures. By Western blot, this antibody detects an ~45 kDa protein from canine striatum representing A2aAR. Immunohistochemical staining of A2aAR in human hippocampus with PA1-042 yields a pattern consistent with plasma membrane staining.
The PA1-042 immunogen is a synthetic peptide corresponding to residues E(373) S H G D M G L P D V E L L S H E L K(391) of canine A2aAR.
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 A2aAR is a glycoprotein that can activate adenylyl cyclase via Gs interactions. Stimulation of A2aAR and the resultant accumulation of cAMP inhibits platelet aggregation and in certain vascular beds is associated with vasodilation and drop in blood pressure. A2aAR is reported to interact with D2 dopamine receptor sites in the brain where they co-express (striatum, olfactory tubercle and nucleus accumbens). Other tissues including heart, kidney, and lung also express A2aAR.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Abnormal calcium handling in atrial fibrillation is linked to up-regulation of adenosine A2A receptors.
PA1-042 was used in western blot to investigate the relationship between calcium signaling and adenosine A receptor level in atrial fibrillation
|Llach A,Molina CE,Prat-Vidal C,Fernandes J,Casadó V,Ciruela F,Lluís C,Franco R,Cinca J,Hove-Madsen L||European heart journal (32:721)||2011|
Adenosine A 2B receptors modulate cAMP levels and induce CREB but not ERK1/2 and p38 phosphorylation in rat skeletal muscle cells.
PA1-042 was used in western blot to investigate the role of adenosine A(2B) on specific signal pathway in rat skeletal muscle cells
|Lynge J,Schulte G,Nordsborg N,Fredholm BB,Hellsten Y||Biochemical and biophysical research communications (307:180)||2003|
Stimulation of A(2A) adenosine receptor phosphorylation by protein kinase C activation: evidence for regulation by multiple protein kinase C isoforms.
PA1-042 was used in western blot to investigate the mechanism of A(2A) adenosine receptor phosphorylation by protein kinase C activation
|Palmer TM,Stiles GL||Biochemistry (38:14833)||1999|