Overview

  • Product name

    Anti-Adenosine Receptor A2a antibody [7F6-G5-A2]
    See all Adenosine Receptor A2a primary antibodies
  • Description

    Mouse monoclonal [7F6-G5-A2] to Adenosine Receptor A2a
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Guinea pig, Hamster, Dog, Non human primates
  • Immunogen

    Full length protein corresponding to Human Adenosine Receptor A2a.
    Database link: P29274

  • Epitope

    Recognises amino acids 213-220 (SQPLPGER) within the third intracellular loop.
  • Positive control

    • This antibody gave a positive result when used in the following methanol fixed cell lines: SKNSH In western blot, this antibody gave a positive result in SHSY-5Y, SK-N-SH, PC12 and human platelet whole cell lysates.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab79714 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 Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 45 kDa).

We recommend using 1% BSA as a blocking agent for western blot.

ICC/IF Use a concentration of 5 µg/ml.

Target

  • Function

    Receptor for adenosine. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase.
  • Sequence similarities

    Belongs to the G-protein coupled receptor 1 family.
  • Domain

    The cytoplasmic C-terminal domain is necessary for targeting the non-ubiquitinated form of this protein to the cell surface.
  • Post-translational
    modifications

    Ubiquitinated. Deubiquitinated by USP4; leading to stabilization and expression at the cell surface.
  • Cellular localization

    Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • A2AAR antibody
    • A2aR antibody
    • AA2AR_HUMAN antibody
    • ADENO antibody
    • Adenosine A2 receptor antibody
    • Adenosine A2a receptor antibody
    • Adenosine receptor A2a antibody
    • Adenosine receptor subtype A2a antibody
    • ADORA 2 antibody
    • ADORA2 antibody
    • ADORA2A antibody
    • hA2aR antibody
    • RDC 8 antibody
    • RDC8 antibody
    see all

Images

  • ICC/IF image of ab79714 stained SKNSH cells. The cells were 100% methanol fixed (5 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab79714 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- mouse (ab150117) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • All lanes : Anti-Adenosine Receptor A2a antibody [7F6-G5-A2] (ab79714) at 5 µg/ml

    Lane 1 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 2 : SK-N-SH (Human neuroblastoma) Whole Cell Lysate
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 4 : Platelet (Human) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 45 kDa


    Exposure time: 2 minutes


    We recommend using 1% BSA as a blocking agent for western blot. Milk is not a suitable blocking agent for this product.

References

This product has been referenced in:

  • Sakano D  et al. Dopamine D2 Receptor-Mediated Regulation of Pancreatic ß Cell Mass. Stem Cell Reports 7:95-109 (2016). Mouse . Read more (PubMed: 27373926) »
  • Niesman IR  et al. Traumatic brain injury enhances neuroinflammation and lesion volume in caveolin deficient mice. J Neuroinflammation 11:39 (2014). WB ; Mouse . Read more (PubMed: 24593993) »
See all 3 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Abreviews
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

Abcam user community

Verified customer

Submitted Jun 10 2013

Answer

Thank you for that reply.

Kate is currently away from the office but I will try to help as best I can with your enquiry. I have checked the two secondary antibodies intended to be used and they seem like they should perform fine with the primary antibodies. As Kate has indicated, I would be happy to provide the alternative antibody ab101678 if your customer would like.

Could you please just confirm I have the correct order number:
xxxx

Then I will arrange for the replacement to be sent to you as soon as possible.

I look forward to receiving your reply.

Read More

Answer

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments.

Before proceeding, could you confirm the secondary antibody used will detect mouse IgG2a isotype? What is the supplier and product number?

If ths antibody is not working with the correct secondary, I would be pleased to arrange an alternative replacement as requested.

I look forward to hearing from you.

Read More

Answer

Thank you for your message and for kindly confirming these details.

I look forward to hearing from you with the results from the samples lysed in RIPA. Please do not hesitate to let me know how you get on. If this still does not work with RIPA, I'll be pleased to arrange a free of charge replacement or credit note.

I hope this will be helpful.

Read More

Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab79714 is tested and covered by our 6 month guarantee for use in WB and human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimize the results. I would also appreciate if you can confirm some further details:

1. I notice the primary antbody ab79714 is a mouse monoclonal IgG2a antibody. The secondary antibody mentioned on the form is goat anti rabbit, which will not detect the mouse primary. I can suggest to use a secondary antibody which is anti mouse IgG, or anti mouse IgG2a.

2. I can recommend to try RIPA lysis buffer to prepare the samples. This may provide amore suitable protein preparation.

3. What were the results from the loading control (GAPDH and b actin)?

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Answer

Thank you for your reply.

I can confirm that I send you the quote for ab79714 in a separate email.

Read More

Answer

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

We like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HepG2)
Specification
HepG2
Fixative
Paraformaldehyde
Permeabilization
Yes - PBS containing 0.2% Triton X100
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Mar 18 2011

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