Recombinant
RabMAb

Recombinant Anti-Adenosine A1 Receptor antibody [EPR6179] (ab124780)

Overview

  • Product name

    Anti-Adenosine A1 Receptor antibody [EPR6179]
    See all Adenosine A1 Receptor primary antibodies
  • Description

    Rabbit monoclonal [EPR6179] to Adenosine A1 Receptor
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues in the extracellular domain of Adenosine A1 Receptor (Human)

  • Positive control

    • WB: Saos 2, SH SY5Y, Caco 2, A549 or 293T cell lysates; IHC-P: Human breast tissue, Mouse cerebrum and hippocampal tissue, Rat cerebellum and cerebrum tissue.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab124780 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
WB 1/1000 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 37 kDa).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/50 - 1/100.

Target

Images

  • All lanes : Anti-Adenosine A1 Receptor antibody [EPR6179] (ab124780) at 1/1000 dilution

    Lane 1 : Saos 2 cell lysate
    Lane 2 : SH SY5Y cell lysate
    Lane 3 : Caco 2 cell lysate
    Lane 4 : A549 cell lysate
    Lane 5 : 293T cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled Goat anti Rabbit at 1/2000 dilution

    Predicted band size: 37 kDa
    Observed band size: 36 kDa
    why is the actual band size different from the predicted?

  • Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling Adenosine A1 with ab124780 at 1/1000 dilution (1.05 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on Purkinje cells in rat cerebellum (PMID: 11137765). The section was incubated with ab124780 for 15 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Adenosine A1 with ab124780 at 1/1000 dilution (1.05 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse cerebrum (PMID: 11137765). The section was incubated with ab124780 for 15 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Overlay histogram showing SH-SY5Y cells stained with ab124780 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab124780, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Adenosine A1 with ab124780 at 1/1000 dilution (1.05 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat cerebrum (PMID: 11137765). The section was incubated with ab124780 for 15 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling Adenosine A1 with ab124780 at 1/1000 dilution (1.05 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse hippocampus (PMID: 11137765, 30315630). The section was incubated with ab124780 for 15 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • ab124780, at 1/100, staining Adenosine A1 Receptor in formalin fixed paraffin embedded Human breast tissue using immunohistochemistry.
  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

ab124780 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for contacting us.
I am sorry to hear that the 2 antibodies do not seem to work as expected.

As we discussed over the phone, ab82477 has not been tested in human and chinese hamster samples. But if you could try it on mouse or rat samples (which are tested species), we could come to a better conclusion regarding the performance of the antibody.

As for ab124780, we did obtain a band for 293T cells and the antibody should give a band for human samples. Would you be able to test 293T cells, also for the RT-PCR signal?

I look forward to hear back from you and assist you further.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Following up my previous message, the immunogen for ab124780, an alternative to ab3460, is also unfortunately unlikely to react with canine Adenosine A1 Receptor. Alignment of the canine receptor amino acid sequence with that of the ab124780 immunogen returns 60% identity. We generally consider this to indicate a low probability of cross-reaction.

Please let me know if we can help in any way.

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Answer

Thank you so much for your reply.

I look forward to hear back from you and wish you good luck with ab124780. This antibodyhas been tested in WB with human samples, and thus, will be guaranteed.

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Answer

I now have some additional information to share with you in regards to the antibodies I discussed in my last email against adenosine receptors.

Anti-Adenosine Receptor A2a

Unfortunately I can confirm that the other two antibodies I suggested against Adenosine Receptor A2a (ab97583 and ab106143) may not be suitable for what you are trying to do. Ab106143 binds in the cytoplasmic region whist the immunogen used to raise ab97583 (residues 255-305) spans the transmembrane, extracellular and cytoplasmic region. It is therefore difficult to tell if ab97583 recognises the extracellular domain but as it is only 8 residues of the whole 50 residue portion, is it unlikely.

Anti-Adenosine Receptor A3

The immunogen used to raisethe ab104502 antibody was taken from the cytoplasmic region of the protein.

I am sorry that I could not bring better news. It seems that the only antibody we can offer which definitely recognises the extracellular region and has been used in flow cytometry previously isagainst the A1 receptor (https://www.abcam.com/Adenosine-A1-Receptor-antibody-EPR6179-ab124780.html).

IF Ican be of any more help please do let me know. I wish you all the best with your research.

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