Overview

  • Product name
    Anti-Adenosine A1 Receptor antibody
    See all Adenosine A1 Receptor primary antibodies
  • Description
    Rabbit polyclonal to Adenosine A1 Receptor
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Rat Adenosine A1 Receptor.

    Read Abcam's proprietary immunogen policy (Peptide available as ab91076.)

  • Positive control
    • This antibody gave a positive signal in Rat Spinal Ganglion Neuronal cell lysate and Mouse EL4 Lymphoma whole cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab82477 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
IHC-P Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

Images

  • All lanes : Anti-Adenosine A1 Receptor antibody (ab82477) at 1 µg/ml

    Lane 1 : Rat Spinal Ganglion Neuronal cell lysate
    Lane 2 : EL4 whole cell lysate (ab7183)

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 36 kDa
    Observed band size: 36 kDa
    Additional bands at: 118 kDa, 18 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute
  • 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.
  • IHC image of ab82477 staining in Rat Adult Brain formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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.

References

This product has been referenced in:
  • Navarro G  et al. Cross-communication between Gi and Gs in a G-protein-coupled receptor heterotetramer guided by a receptor C-terminal domain. BMC Biol 16:24 (2018). Read more (PubMed: 29486745) »
  • Zhai W  et al. A1 adenosine receptor attenuates intracerebral hemorrhage-induced secondary brain injury in rats by activating the P38-MAPKAP2-Hsp27 pathway. Mol Brain 9:66 (2016). IF ; Rat . Read more (PubMed: 27301321) »
See all 9 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA
Sample
Human Tissue sections (Brain cortex)
Specification
Brain cortex
Permeabilization
Yes - TBS-T (0.05% tween)
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 28 2014

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

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.

Read More

Answer

Thank you for submitting an Abreview of ab82477. As you may have noticed, your review has now been published on our website.

Since you obtained poor results using the antibody in a untested species, we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.

Based on the information in your review, I feel that altering the following step in your protocol may help improve your results:

- try the primary antibody on mouse or rat samples as positive control
- load more human protein
- check the secondary antibody

If you decide to follow these suggestions, please let me know if they are helpful.

I look forward to your reply.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-20% or 10%)
Sample
Human Cell lysate - whole cell (CHO-cells)
Specification
CHO-cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Mr. David Wilkes

Verified customer

Submitted May 11 2012

Answer

Thank you very much for your interest in ab82477.

This antibody was tested in IHC-P from our lab using Leica BondTM system with the standard protocol F. If you do not have access to this system I suggest to use a standard protocol that can be found here:
https://www.abcam.com/index.html?pageconfig=resource&rid=11384

https://www.abcam.com/index.html?pageconfig=resource&rid=13046

The antigen retrieval used to generate the image on the datasheet was Citrate buffer pH6 for 20min. I suggest to incubate the antibody overnight at 4C to achieve the best saturation.

To our knowledge, ab82477 has not been tested in IHC-Fr. Therefore, I can offer a discount off a future purchase if you buyab82477 now, test it in IHC-Fr and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody or the value of your next order..

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and testab82477 in IHC-Fr. I will then send a discount code. This code must be issued before purchasing ab82477 so please wait for my reply before ordering.

2. Purchase ab82477 either by phone, fax, or online (www.abcam.com).

3. Test it in IHC-Fr.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyproduct ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab82477 turns out to be unsuitable for IHC-Fr, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Question
Answer

DISCOUNT CODE: ***
Expiration date: August 4th, 2012

I am very pleased to hear you would like to accept our offer and test ab82477 on human samples. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for human and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for your telephone enquiry this morning.

As discussed on the telephone, I can confirm we do have a Adenosine A1 Receptor antibody tested in rat and IHC-P:

ab82477 Adenosine A1receptor antibody

However, I am sorry that to our knowledge, ab82477 and our other Adenosine A1 receptor antibodies have not been tested in IHC-Fr yet.

Therefore, I can offer a discount off a future purchase if you buy a82477b now, test it in IHC-Fr and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab82477 in IHC-Fr. I will then send a discount code. This code must be issued before purchasing ab82477 so please wait for my reply before ordering.

2. Purchase ab82477 either by phone, fax, or online (www.abcam.com).

3. Test it in IHC-Fr.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab82477 turns out to be unsuitable for IHC-Fr, you will still receive the discount on your next purchase after your Abreview has been submitted.

With regards to the protocol, this particular antibody is tested using an automated system, and so the protocol would not be relevant to other laboratories. I can suggest to review our immunostaining protocols from the protocols page. This provides protocol guidelines, however we recommend that procedures will often require individual optimization by the end user:
https://www.abcam.com/index.html?pageconfig=popular_protocols

Please let me know if you have any questions about the protocol or theoffer and I would be happy to help you further.

The Terms and Conditions of the testingoffer can be found at: www.abcam.com/collaborationdiscount.

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Answer

I attached the WB for ab82477. The first lane is rat spinal ganglion neuronal cell lysate. The second lane is the positive control- ab7183. Both antibody westerns can be found on the datasheet on the abcam website. Good luck.

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1-10 of 11 Abreviews or Q&A

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