Anti-Adenosine A1 Receptor antibody

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adenosine A1 Receptor antibody (AB82477)

IHC image of ab82477 staining in Rat Adult Brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab82477, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Adenosine A1 Receptor antibody (AB82477)

ICC/IF image of ab82477 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab82477, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) PC12 cells at 5µg/ml.

• WB

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Western blot - Anti-Adenosine A1 Receptor antibody (AB82477)

All lanes:

Western blot - Anti-Adenosine A1 Receptor antibody (ab82477) at 1 µg/mL

Lane 1:

Rat Spinal Ganglion Neuronal cell lysate at 10 µg

Lane 2:

Western blot - EL4 whole cell lysate (<a href='/en-us/products/cell-lysates/el4-whole-cell-lysate-ab7183'>ab7183</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 37 kDa

Observed band size: 118 kDa,18 kDa,36 kDa

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Exposure time: 1min

• WB

CiteAb

Western blot - Anti-Adenosine A1 Receptor antibody (AB82477)

Adenosine A1 Receptor western blot using anti-Adenosine A1 Receptor antibody ab82477. Publication image and figure legend from Zhai, W., Chen, D., et al., 2016, Mol Brain, PubMed 27301321.

ab82477 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab82477 please see the product overview.

Changes in adenosine receptor expression in vivo and in vitro. a Western blot analysis of adenosine receptor expression at different time points following ICH in vivo. A1AR levels peaked at 48 h. b Quantification of the results in panel A. Bars represent relative protein levels. The mean values of the protein levels in the sham group were normalized to 1.0. *p < 0.05 for the 48 h ICH group versus the sham group. c Western blots of adenosine receptor expression in vitro. A1AR levels peaked at 48 h. d Quantification of the results in panel C. Bars represents relative protein levels. The mean values of the protein levels in the control group were normalized to 1.0. *p < 0.05 for the 48 h OxyHb group versus the control group. e and f Immunofluorescence in vivo. Double immunofluorescence analysis was performed with A1AR antibodies (green) and neuronal or astrocyte marker (NeuN/GFAP, red), and nuclei were fluorescently labeled with DAPI (blue). Scale bar = 32 μm. G : Immunofluorescence in vitro. Double immunofluorescence analysis was performed with A1AR antibodies (green) and neuronal marker (NeuN, red), and nuclei were fluorescently labeled with DAPI (blue). Scale bar = 20 μm

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