This is a knockout-validated antibody summary, based on the publication "Astrocytic adenosine receptor A2A and Gs-coupled signaling regulate memory", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. (a) Representative photomicrographs of striatal and cortical sections from 12–13-month-old GFAP-Cre/loxP-Adora2a mice and 2-month-old global homozygous A2A receptor knockout mice (A2A-gKO) immunostained for the A2A receptor. Insets (i–iv): magnified views of the boxed regions. Scale bar: 500 µm. n = 3 mice per genotype. (b–c) Representative confocal micrographs (top) and quantitative histological analyses (bottom) of hippocampal CA1 sections from 12–13-month-old wild-type controls (Control) and A2A-cKO mice, and 2-month-old A2A-gKO mice co-immunostained for the A2A receptor (red) and an astrocyte marker (GFAP, green) or a neuronal marker (NeuN). Cell nuclei were labeled with DAPI (blue). Scale bar: 10 µm. From [1].

Mouse monoclonal IgG2ak

Company: Millipore, now MilliporeSigma

Antibody: A2A

Catalog number: 05-717

Summary: Mouse monoclonal IgG2ak raised against 6His-tagged full-length human recombinant A2A receptor. Epitope mapped to SQPLPGER in the third intracellular loop. Reacts with human, rat, mouse, rabbit and dog. Suitable for western blotting, immunohistochemistry and flow cytometry.

Western blot

brain sections from GFAP-Cre/loxP-Adora2a mice and global homozygous A2A receptor knockout mice

blocking reagent provided in the mouse-on-mouse kit (Vector Laboratories)

striatal and cortical sections: 1:200 overnight at room temperature

hippocampal sections: 1:10,000 overnight at 4°C

striatal and cortical sections: Donkey anti-mouse secondary antibodies conjugated to Alexa fluorophores (Life Technologies, 1:500, A21202 and A21203)

hippocampal sections: Tyramide Signal Amplification™- Direct (Red) system (1:100, NEN Life Sciences, Boston, MA)

striatal and cortical sections: Nikon TE300 microscope and Spot RT digital CCD camera and imaging software (Diagnostic Instruments).

hippocampal sections: Zeiss 63X (N.A. 1.4) objective on an Axiovert 35 microscope (Zeiss) with an attached MRC1024 laser scanning confocal microscope system (BioRad)

The same clone (7F6-G5-A2) is sold as Novus Biologicals NBP1-39474; Santa Cruz Biotechnology sc-32261; Abcam ab115250; EMD Millipore 05-717.