Overview

  • Product name

    Anti-Renin Receptor antibody
    See all Renin Receptor primary antibodies
  • Description

    Rabbit polyclonal to Renin Receptor
  • Host species

    Rabbit
  • Specificity

    Detects endogenous levels of total Renin Receptor protein.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, ELISAmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide corresponding to Human Renin Receptor (internal sequence).

  • Positive control

    • Human brain tissue COLO205 and HeLa cell extracts HeLa cells
  • General notes

    Protein target previously labeled as POU3F2.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride

    Without Mg2+ and Ca2+
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab64957 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 1/500 - 1/1000. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa).
IHC-P 1/50 - 1/100.
ICC/IF 1/500 - 1/1000.
ELISA 1/20000.

Target

  • Function

    Functions as a renin and prorenin cellular receptor. May mediate renin-dependent cellular responses by activating ERK1 and ERK2. By increasing the catalytic efficiency of renin in AGT/angiotensinogen conversion to angiotensin I, it may also play a role in the renin-angiotensin system (RAS).
  • Tissue specificity

    Expressed in brain, heart, placenta, liver, kidney and pancreas. Barely detectable in lung and skeletal muscles. In the kidney cortex it is restricted to the mesangium of glomeruli. In the coronary and kidney artery it is expressed in the subendothelium, associated to smooth muscles where it colocalizes with REN. Expressed in vascular structures and by syncytiotrophoblast cells in the mature fetal placenta.
  • Involvement in disease

    Defects in ATP6AP2 are a cause of mental retardation X-linked with epilepsy (MRXE) [MIM:300423]. MRXE is a syndromic mental retardation. Patients manifest mild to moderate mental retardation associated with epilepsy, delays in motor milestones and speech acquisition in infancy.
  • Post-translational
    modifications

    Phosphorylated.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • APT6M8 9 antibody
    • APT6M8-9 antibody
    • ATP6AP2 antibody
    • ATP6IP2 antibody
    • ATP6M8-9 antibody
    • ATPase H(+)-transporting lysosomal accessory protein 2 antibody
    • ATPase H(+)-transporting lysosomal-interacting protein 2 antibody
    • ATPase H+ transporting lysosomal accessory protein 2 antibody
    • ATPase H+ transporting lysosomal interacting protein 2 antibody
    • ATPase H+ transporting lysosomal vacuolar proton pump membrane sector associated protein M8 9 antibody
    • ATPase membrane sector associated protein M8 9 antibody
    • ATPase, H+ transporting, lysosomal (vacuolar proton pump) membrane sector associated protein M8 9 antibody
    • CAPER antibody
    • ELDF10 antibody
    • Embryonic liver differentiation factor 10 antibody
    • ER localized type I transmembrane adaptor antibody
    • ER-localized type I transmembrane adaptor antibody
    • HT028 antibody
    • M8 9 antibody
    • M8-9 antibody
    • MGC99577 antibody
    • MRXE antibody
    • MSTP009 antibody
    • N14F antibody
    • Renin receptor antibody
    • Renin/prorenin receptor antibody
    • RENR_HUMAN antibody
    • V ATPase M8 9 subunit antibody
    • V ATPase M8.9 subunit antibody
    • V-ATPase M8.9 subunit antibody
    • Vacuolar ATP synthase membrane sector associated protein M8 9 antibody
    • Vacuolar ATP synthase membrane sector-associated protein M8-9 antibody
    • vacuolar proton ATP synthase membrane sector associated protein M8 9 antibody
    • XMRE antibody
    see all

Images

  • All lanes : Anti-Renin Receptor antibody (ab64957) at 1/500 dilution

    Lane 1 : COLO205 cell extract
    Lane 2 : HeLa cell extract
    Lane 3 : HeLa cell extract with immunising peptide at 5 µg

    Lysates/proteins at 5 µg per lane.

    Predicted band size: 39 kDa
    Observed band size: 39 kDa

  • Immunohistochemical analysis of paraffin-embedded human brain tissue using ab64957 at 1/50 dilution. Right hand panel was also treated with immunising peptide.
  • Immunofluorescence analysis of HeLa cells, using ab64957 at 1/500 dilution. Right hand panel was also treated with immunising peptide.

References

This product has been referenced in:

  • Alzayadneh EM & Chappell MC Nuclear expression of renin-angiotensin system components in NRK-52E renal epithelial cells. J Renin Angiotensin Aldosterone Syst N/A:N/A (2014). ICC/IF . Read more (PubMed: 24961503) »
  • White AJ  et al. Expression of classical components of the renin-angiotensin system in the human eye. J Renin Angiotensin Aldosterone Syst N/A:N/A (2014). IHC ; Human . Read more (PubMed: 25287897) »
See all 5 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate, pH6
Permeabilization
No
Specification
Brain
Blocking step
Dako dual enzyme block, Dako protein block as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 21°C
Fixative
Formaldehyde

Dr. Jean-Martin Lapointe

Verified customer

Submitted Apr 21 2015

Question
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 and it is regrettable the results have not been succesful.

I am sorry to confirm that the order number relates to an order placed 15th Dec 2011, which shipped 5th January. Therefore,this item was purchase 8 months ago and so is regrettably past the 6 month guarantee period. I am pleased to make an exception in this case offer a free of charge replacement or credit note in compensation as a gesture of goodwill. However, please note we would encourage customers to contact us as soon as possible if they encounter any difficulties, and within the 6 months of purchase so that it is still within guarantee.

Thank you for your understanding of our guarnatee policy and for your help and cooperation in this case. I look forward to hearing from you with details of how the customer would like to proceed.

Read More

Question

We have received a response from our customer;

As explained, I'm rather disappointed with Abcam’s response and original lack of foresight to tell us that the original antibody was unproven. I also think that it us reasonable to expect us to test the antibody in human samples with upregulated renin-prorenin receptors which are not exactly easy to come across

Furthermore after reading one of the two reference articles provided by Abcam on their Website, I am still uncertain about the IHC use of other antibody Ab40790. The only rat paper to use it Huang & Siragy 2009 Endocrinology actually claim it came from Santa Cruz in one section of their paper or as a personal gift from another researcher in a second section.

The immunofluorescence photographs on the web site of kidney tubules show an awful lot of background fluorescence with no clear confirmation of actual localised staining.

All very confusing.

I will only try the Ab40790 if Abcam positively guarantee its use in rat kidney and brain.

We intend to use it as a immunohistochemical applical with DAB staining

Species will be rat (ren-2 over expressing transgenic rat versus Sprague Dawley controls)

Samples: Kidney paraffin embedded 10% nBF fixed with microwave antigen retrieval 5 min.

Brain cryosections post fixed in acetone or NBF.

Can you please ask ABCAM if they will provide us with a test sample.

Is there anyway we can help our customer further, by providing him with a FOCR sample of AB40790, to keep him happy.

Let us know your thoughts and we hope to hear from you soon.

Kind regards,

Read More
Answer

Thank you for your message.

I am sincerely sorry, I have gone back through the details and realize that I originally made a mistake regarding tested species. It is mouse, chicken and cow that this antibody isonly predicted for. The antibody is indeed tested and covered by our guarantee for rat. I fully understand the customers concerns regarding this issue, please pass on my apologies forthe confusion.

In this case, as rat is a species we can cover by the guarantee, I would appreciate if they could consider trying the suggestions made in the previous email. In order to help with our investigation of this case before deciding how to proceed, I would appreciate also if you are able to provide more detail as requested in the previous email. This information will also be helpful to our quality monitoring.

1. Could you confirm which buffer was used for antigen retrieval? Were different time points tried (for example 2, 5. 10 and 20 minutes)? Antigen retrieval can sometimes require some optimization.

2. How long was the permeabilization? We recommend 10 minutes only.

3. We recommend not to mix blocking agents in an experiment. Try 5% serum only for the blocking step, for 1 hour. this should help to prevent over - blocking. Try 2% serum in the antibody dilution buffers.

4. Is the secondary antibody working well with other primary antibodies?

5. Please confirm the order number and date of purchase.

With regards to receiving a sample vial of alternative ab40790, I am sorry to confirm that because we sell over 90 000 products, it is regrettably not possible for us to store and send sample vials.I will be pleased to provide a free of charge replacement or credit note if the ab64957 antibody is not working with the suggestions provided and the item has been purchased in the last 6 months. Or an alternative replacement if this is what the customer prefers.

With regards to the publications for ab40790, we do produce many antibodies in house, howeverwe also source many externally. Those that are sourced externally are often also sold by other companies. Therefore, the publications will be from the same antibody.

Thank you for your time. I look forward to hearing from you with the requested information and hope we can resolve this case as soon as possible.

Read More

Question

Product code: 64957
Lot number: GR40186-1

Inquiry:
Antibody Storage Conditions (temperature/reconstitution etc) -20 C freezer
Description of the Problem (high background, low signal, non-specific staining etc) No staining at all
Sample (Species/Tissue/Cell Type/Cell Line etc)
REN-2 over expressing transgenic rat Cryo sections from unfixed Rat Brain and Paraffin Embedded 10% NBF fixed Rat Kidney
Fixation of Sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration) -20 C Acetone fixation for brain cryosections No further processing with kidney
Antigen Retrieval (Enzymatic method, Heat mediated technique etc)
No antigen retrieval performed on rat brain
Paraffin Embedded 10% NBF fixed Rat Kidney sections subjected to microwave antigen retrieval
Permeabilization step 0.3% Triton X-100
Blocking Conditions (Buffer/time period, Blocking agent) PBS in 0.1% BSA and 10% goat serum 2hr
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step)
1-20-1/100 Abcam rabbit polyclonal in TBS with BSA ; 0.3% triton X overnight at 4C
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step)
1/100-1/500 Pierce Goat anti-rabbit HRP conjugated in 0.1% BSA with TBS
Detection Method DAB detection (Vector Scientific) Immunohistochemistry
Positive and negative controls used (please specify) -ve control conducted using no primary Ab
OPTIMIZATION ATTEMPTS (PROBLEM SOLVING) How many times have you tried the IHC? 5 times with both brain and kidney
Have you run a “No Primary” control? Yes
Do you obtain the same results every time? Yes What steps have you altered? Blocking, fixation, dilution and DAB steps all proved negative.
Additional Notes:

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 willgive us with vital information for our monitoring of product quality. I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forIHC-P (paraffin embedded)and human samples.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there few tips to provide on this occasion to help improve the results.

However, I can recommend you may like to consider the following:

1. I am sorry to confirm that this antibody is not tested and covered by our 6 month guarantee for rat samples. This is predicted only due to high sequence alignment. I am sorry this regrettably means we are not able to provide credit or replacement in this case for rat. I can recommend to consider including a positive controlsample fromthe tested species, human. We can provide our guarantee for human samples.

2. Could you confirm which buffer was used for antigen retrieval? Were different time points tried (for example 2, 5. 10 and 20 minutes)? Antigen retrieval can sometimes require some optimization.

3. How long was the permeabilization? We recommend 10 minutes only.

4. We recommend not to mix blocking agents in an experiment. Try 5% serum only for the blocking step, for 1 hour. this should help to prevent over - blocking. Try 2% serum in the antibody dilution buffers.

5. Is the secondary antibody working well with other primary antibodies?

6. Please confirm the order number and date of purchase.

Thank you for your cooperation. I look forward to hearing from you withthe further requesteddetails and results from humanpositive control samples.

Read More

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