Anti-Adenosine Receptor A2a antibody (ab3461)

Overview

  • Product nameAnti-Adenosine Receptor A2a antibody
    See all Adenosine Receptor A2a primary antibodies
  • Description
    Rabbit polyclonal to Adenosine Receptor A2a
  • SpecificityDetects adenosine receptor A2a. This antibody does not detect other AR subtypes.
  • Tested applicationsSuitable for: ICC, WB, IHC-P, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
    Predicted to work with: Horse
  • Immunogen

    Synthetic peptide corresponding to Dog Adenosine Receptor A2a aa 373-391.
    Sequence:

    ESHGDMGLPDVELLSHELK

Properties

Applications

Our Abpromise guarantee covers the use of ab3461 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC 1/10 - 1/100.
WB 1/1000.
IHC-P 1/20 - 1/200.
IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 10 µg/ml.

Target

Anti-Adenosine Receptor A2a antibody images

  • 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 ºC 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.

  • 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.

  • 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.

  • 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.

  • Anti-Adenosine Receptor A2a antibody (ab3461) at 1/1000 dilution + Human heart tissue lysate at 20 µg

    Secondary
    Donkey HRP-conjugate anti-rabbit at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 3 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • All lanes : Anti-Adenosine Receptor A2a antibody (ab3461) at 1/500 dilution

    Lane 1 : Human placenta cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Mouse liver cell lysate

    Lysates/proteins at 25 µg per lane.
  • 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.

References for Anti-Adenosine Receptor A2a antibody (ab3461)

This product has been referenced in:
  • Munson MJ  et al. mTOR activates the VPS34-UVRAG complex to regulate autolysosomal tubulation and cell survival. EMBO J 34:2272-90 (2015). Read more (PubMed: 26139536) »
  • Oláh A  et al. Cannabidiol exerts sebostatic and antiinflammatory effects on human sebocytes. J Clin Invest 124:3713-24 (2014). ICC/IF ; Human . Read more (PubMed: 25061872) »

See all 4 Publications for this product

Product Wall

Application ELISA
Sample Human Cell (Cell Membrane)
Specification Cell Membrane
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Type Sandwich (Detection)
Username

Tammy Zhai

Verified customer

Submitted Apr 22 2015

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (8)
Sample Human Tissue lysate - whole (Heart)
Specification Heart
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 10 2013

Hi Daniela, I did an alignment between the rat sequence and the immnunogen sequence for ab3461 and I would not recommend using this antibody in rat. The level of similarity between the two is to low. I would however recommend ab101678, it has not been ...

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