Recombinant
RabMAb

Recombinant Anti-Angiotensin II Type 2 Receptor antibody [EPR3876] (ab92445)

Rabbit recombinant monoclonal Angiotensin II Type 2 Receptor antibody [EPR3876]. Validated in WB, IP and tested in Mouse, Rat, Human. Cited in 14 publication(s). Independently reviewed in 1 review(s).

Overview

  • Product name

    Anti-Angiotensin II Type 2 Receptor antibody [EPR3876]
    See all Angiotensin II Type 2 Receptor primary antibodies
  • Description

    Rabbit monoclonal [EPR3876] to Angiotensin II Type 2 Receptor
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt,ICC or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Angiotensin II Type 2 Receptor aa 300-400 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HepG2, Human heart, Mouse heart, and Rat heart lysates. IP: Mouse heart cells.
  • General notes

     

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab92445 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. Predicted molecular weight: 41 kDa.
IP 1/10 - 1/100.
  • Application notes
    Is unsuitable for Flow Cyt,ICC or IHC-P.
  • Target

    Images

    • All lanes : Anti-Angiotensin II Type 2 Receptor antibody [EPR3876] (ab92445) at 1/1000 dilution (Purified)

      Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
      Lane 2 : Human heart lysates
      Lane 3 : Mouse heart lysates
      Lane 4 : Rat heart lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 41 kDa
      Observed band size: 41 kDa

    • ab92445 (purified ) at 1/20 dilution (0.2ug) immunoprecipitating Angiotensin II Type 2 Receptor in Mouse heart lysate .
      Lane 1 (input): Mouse heart lysate 10ug
      Lane 2 (+): ab92445 & Mouse heart lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92445 in Mouse heart lysate
      For western blotting, VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    References

    This product has been referenced in:

    See all 17 Publications for this product

    Customer reviews and Q&As

    1-10 of 14 Abreviews or Q&A

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (heart)
    Gel Running Conditions
    Non-reduced Denaturing (12%)
    Loading amount
    30 µg
    Specification
    heart
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Mar 01 2019

    Application
    Western blot
    Sample
    Pig Tissue lysate - whole (Newborn kidney)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    40 µg
    Specification
    Newborn kidney
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Dec 28 2017

    Answer


    AT2R is a 41 kDa protein that contains a large number of potential glycosylation sites, and depending on the amount of post-translational glycosylation, the protein may run at a variety of sizes between 41 and 58 kDa. The SwissProt link below depicts the glycosylation sites:
    http://www.uniprot.org/uniprot/P50052
    Additionally, in viewing the western blot images associated with our other AT2R antibody that has been tested in western blot (ab19134), it seems the protein does run at a variety of sizes depending on the amount of glycosylation:
    ab19134 western blot image: https://www.abcam.com/Angiotensin-II-Type-2-Receptor-antibody-ab19134-reviews.html?intabreviewid=7301
    Thus, it seems that both ab92445 and ab78747 are specifically detecting AT2R, with varying molecular weights depending on the amount of glycosylation.

    Read More

    Answer

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

    I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forWB and in Mouse, Rat andHuman samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

    I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

    I would appreciate if you could also provide an image which would help us to assess the results, including the endogenous negative and positive controls.

    As discussed on the telephone, if the endogenous negative controls are giving very low signal, then I can suggest it would be beneficial to consider revewing the siRNA knock out procedure. This following page from our website provides examples of checks and controls for siRNA experiments, including assessing the results and troubleshooting. I hope you will find this useful:

    https://www.abcam.com/index.html?pageconfig=resource&rid=12137


    Thank you for your time and cooperation. We look forward to receiving the completed questionaire.




    Order Details
    Antibody code:

    Problem
    Choose: Non-specific band Multiple bands No signal or weak signal High background

    Lot number

    Purchase order number
    or preferably Abcam order number:



    General Information
    Antibody storage conditions (temperature/reconstitution etc)


    Description of the problem (high background, wrong band size, more bands, no band etc.)


    Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


    Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


    Amount of protein loaded


    Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


    Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


    Detection method (ECL, ECLPlus etc.)


    Positive and negative controls used (please specify)



    Optimization attempts (problem solving)
    How many times have you tried the Western?



    Have you run a "No Primary" control?
    Yes No

    Do you obtain the same results every time?
    Yes No
    e.g. are the background bands always in the same place?


    What steps have you altered?


    Additional Notes:


    Image:
    We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

    Read More

    Question
    Answer

    According to our records, 92445 Anti-Angiotensin II Type 2 Receptor antibody [EPR3876] was proving difficult to use inWB and we were in contact in order to help resolve the issue.

    Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

    If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

    We wish you the best of luck with your research and look forward to a reply.

    Read More

    Question
    Answer

    Yes of course, thanks for your reply.

    We don't have any specific recommendations for western blotting protocol for ab32212. Although a good starting dilution for western blotting using ab32212 is noted as 1:5000.

    ab52174 is antibody against a phosphorylated epitope so with this antibody it is best to block with BSA for sure instead of milk, as milk contains phosphorylated proteins which may bind the antibody, creating a high background. A recommended starting concentration for this antibody is 1:1000.

    I hope this is helpful. Please contact me again if you have any further questions.

    Read More

    Answer

    Thanks for your email.

    I'm sorry but I don't quite understand your question. What is it that you requested for ab32213 and ab52174?

    Thanks for the additional details.

    Read More

    Answer

    Thank you for your enquiry.

    i have checked and none ofthese 3 antibodies have a specific western blot protocol associated with them, although typically a 1:1000 dilution, or 1ug/ml primaryantibody concentration works well as a starting point for western blotting.

    A good blocking buffer we usually recommend is 5% BSA in TBST.

    A good antibody diluent to use is 1% BSA in TBST.

    Which antibody are you using to stain the rat mesenteric artery extracts?

    Read More

    Answer

    Thanks for your email.

    For ab11734 I'd recommend starting with a dilution of 1: 250 and dilute ab87436 to 1:1000. The primary antibody diluent could be TBS with 0.05% Tween 20.

    I hope this is helpful. Please contact me again if you have any further questions.

    Read More

    Answer

    Thank you for your enquiry.

    Here is an antibody that has be used in western blotting and IHC on rat samples for ACE 1:
    https://www.abcam.com/Angiotensin-Converting-Enzyme-1-antibody-9B9-ab77990.html

    Here is an antibody that has be used in western blotting and IHC on rat samples for ACE 2:
    https://www.abcam.com/Angiotensin-Converting-Enzyme-2-antibody-EPR4435-2-ab108252.html

    I hope this is helpful. Please contact me again if you have any further questions.

    Read More

    1-10 of 14 Abreviews or Q&A

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