Anti-Adenosine Receptor A2a antibody

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adenosine Receptor A2a antibody (AB3461)

ab3461 labelling Adenosine Receptor A2a in the cytoplasm and membrane of Mouse testis tissue (right) compared with a negative control (left) by Immunohistochemisty (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tissue sections were incubated with the primary antibody (1 : 20 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit IgG was as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

• ICC

Lab

Immunocytochemistry - Anti-Adenosine Receptor A2a antibody (AB3461)

ICC/IF image of ab3461 stained SKNSH 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 (ab3461, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adenosine Receptor A2a antibody (AB3461)

ab3461 labelling Adenosine Receptor A2a in the cytoplasm and membrane of Human testis tissue (right) compared with a negative control (left) by Immunohistochemisty (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tissue sections were incubated with the primary antibody (1 : 200 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit IgG was as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

• ICC

Supplier Data

Immunocytochemistry - Anti-Adenosine Receptor A2a antibody (AB3461)

Immunocytochemistry/Immunofluorescence analysis of Adenosine Receptor A2a (green) showing staining in the cytoplasm of U251 cells (right) compared to a negative control (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 incubated with ab3461 in 3% BSA-PBS at a dilution of 1 : 20 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 fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adenosine Receptor A2a antibody (AB3461)

ab3461 labelling Adenosine Receptor A2a in the cytoplasm and membrane of Human placenta tissue (right) compared with a negative control (left) by Immunohistochemisty (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tissue sections were incubated with the primary antibody (1 : 100 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit IgG was as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

• WB

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Western blot - Anti-Adenosine Receptor A2a antibody (AB3461)

All lanes:

Western blot - Anti-Adenosine Receptor A2a antibody (ab3461) at 1/500 dilution

Lane 1:

Human placenta cell lysate at 25 µg

Lane 2:

HepG2 cell lysate at 25 µg

Lane 3:

HeLa cell lysate at 25 µg

Lane 4:

Mouse liver cell lysate at 25 µg

Predicted band size: 44 kDa

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