Anti-Natriuretic Peptide Receptor C antibody (ab37617)

Overview

  • Product name
    Anti-Natriuretic Peptide Receptor C antibody
    See all Natriuretic Peptide Receptor C primary antibodies
  • Description
    Rabbit polyclonal to Natriuretic Peptide Receptor C
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, ELISA, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human Natriuretic Peptide Receptor C aa 67-97 (N terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).

  • Positive control
    • WB: HL-60 cell lysate IHC-P: human breast carcinoma and kidney tissues. ICC/IF: HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab37617 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
ICC/IF 1/200.
ELISA 1/1000.
WB 1/100 - 1/500. Detects a band of approximately 63 kDa (predicted molecular weight: 60 kDa).
IHC-P 1/50 - 1/100.

Target

  • Function
    Receptor for the natriuretic peptide hormones, binding with similar affinities atrial natriuretic peptide NPPA/ANP, brain natriuretic peptide NPPB/BNP, and C-type natriuretic peptide NPPC/CNP. May function as a clearance receptor for NPPA, NPPB and NPPC, regulating their local concentrations and effects. May regulate diuresis, blood pressure and skeletal development. Does not have guanylate cyclase activity.
  • Sequence similarities
    Belongs to the ANF receptor family.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ANP C antibody
    • ANP-C antibody
    • ANPC antibody
    • ANPR-C antibody
    • ANPRC antibody
    • ANPRC_HUMAN antibody
    • Atrial natriuretic peptide C type receptor antibody
    • Atrial natriuretic peptide clearance receptor antibody
    • Atrial natriuretic peptide receptor 3 antibody
    • Atrial natriuretic peptide receptor type C antibody
    • Atrionatriuretic peptide receptor C antibody
    • C5orf23 antibody
    • FLJ14054 antibody
    • GUCY2B antibody
    • MGC22189 antibody
    • Natriuretic peptide receptor C/guanylate cyclase C (atrionatriuretic peptide receptor C) antibody
    • Natriuretic peptide receptor C/guanylate cyclase C antibody
    • NPR 3 antibody
    • NPR C antibody
    • NPR-C antibody
    • Npr3 antibody
    • NPRC antibody
    see all

Images

  • Anti-Natriuretic Peptide Receptor C antibody (ab37617) at 1/1000 dilution + HL-60 cell lysate at 35 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/5000 dilution

    Predicted band size: 60 kDa



    Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling Natriuretic Peptide Receptor C with ab37617. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at 38°C; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A peroxidase-conjugated Goat anti-rabbit polyclonal (ready to use) was used as the secondary antibody.

  • ab37617 staining Natriuretic Peptide Receptor C in the Human HeLa cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X-100 in PBS  for 30 minutes. Samples were incubated with primary antibody (1/200) for 2 hours. An Alexa Fluor®488-conjugated Donkey anti-rabbit (1/1000) was used as the secondary antibody.

  • Anti-Natriuretic Peptide Receptor C antibody (ab37617) at 1/1000 dilution + HL-60 cell lysate at 35 µg/ml

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/5000 dilution

    Developed using the ECL technique.

    Predicted band size: 60 kDa
    Observed band size: 63 kDa
    why is the actual band size different from the predicted?



    Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.

  • ab37617, at a 1/50 dliution staining human breast carcinoma by immunohistochemistry, formalin fixed paraffin embedded tissue, followed by AEC staining.

References

This product has been referenced in:
  • Gu Y  et al. Aberrant pro-atrial natriuretic peptide/corin/natriuretic peptide receptor signaling is present in maternal vascular endothelium in preeclampsia. Pregnancy Hypertens 11:1-6 (2018). IHC-P . Read more (PubMed: 29523263) »
  • Zhao F  et al. Molecular subgroups of adult medulloblastoma: a long-term single-institution study. Neuro Oncol 18:982-90 (2016). IHC-P ; Human . Read more (PubMed: 27106407) »
See all 7 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (HUVEC)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
HUVEC
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 11 2018

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Sample
Mouse Tissue lysate - whole (Liver)
Specification
Liver
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 23 2014

Answer

Thank you very much for providing that information. It has been very helpful in understanding the results you have seen so far in your experiment.

I am sorry but Kate is currently away from the office but I will try my best to help you with these problems.

It is clear that you have put a lot of effort in to try and optimise the results with these antibodies. It is regrettable that the results so far have not been as hoped. Would you mind clarifying a few further details for me in order to more fully understand what may be contributing to this?

1. You have mentioned that you have purchased three antibodies from us. Is the third one working as expected? Which antibody was this?

2. Have you been performing any staining of the tissues with any other antibodies? How has this staining performed?

3. How were the tissues fixed (time and temperature in formalin)?

4. Can I ask, the antigen retrieval, am I correct in thinking you tried the three different buffers individually, each for 20 minutes at 97oC? How was this performed, in a microwave or pressure cooker etc? You also mentioned it being performed for 30 minutes, 1 hour, 2 hours and overnight, could you clarify this?

Once I have this information I hope to be able to understand more fully what may be contributing to the staining observed currently.

look forward to receiving your reply.

Read More

Question

Thanks for your attention.

I am using human breast carcinoma tissue as positive control for NPR3 antibody and skin (keratinocytes) as positive control for SFRP1 antibody (like descripted in your datasheet). I started my reactions with 1:200 concentration for NPR3 and 1:2000 for SFRP1, as suggested by Michael Taylor, MD in his paper "Medulloblastoma comprises four distinct molecular variants".

However, after five differents attempts (over night, 60 minutes and 1 hour of incubation, with Ultra Vision Plus detection (Thermo)), I think this protocols worked better: 1:50 concentration for NPR3 with 2 hours of incubation, pH 6,0, antigen retrieval in PT-Link (DAKO) and DAKO detection.

In case of SFRP1, I did the same protocols above with up 1:500 in concentration and I didn't get any staining.

Please find the best images attached.

Best regards.


2012/10/12 <mailto:technical@abcam.com>














https://www.abcam.com/index.html?utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Header








Dear Gisele



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

I would like to reassure you that our antibodies are tested and covered by our 6 month guarantee for the tested species and applications listed on the individual datasheets. 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 would appreciate if you could let me know which application you have been using. Also which type of samples you have tested. Once I know which application you are using, I can obtain the protocol from the laboratory and will be pleased to send this on to you.

I would also appreciate if you are able to provide an image which would help us to assess the results.

Thank you for your time and cooperation. I look forward to hearing from you with the requested information.

Help us improve our service.
https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=4242373


Best regards,
Kate

Kate Hayes
Scientific Support Supervisor
Abcam plc
https://www.abcam.com/


Your original inquiry to Abcam:














From: Gisele Caravina de Almeida [mailto:giselecaravina@gmail.com]
Sent: 11 October 2012 16:22
To: Licensing; Careers; Webmaster; Gisele Caravina de Almeida
Subject: I need help about my reactions to SFRP1 and NPR3









Hello,







My name is Gisele and I bought three antibodies of Abcam but two of them aren´t working: SFRP1 (ab4193) and NPR3 (ab37617). I did several protocols but I haven´t got satisfactory result.



I would like to get your protocols for these antibodies to repeat my reactions.



Thanks.










Abcam Customer Services and Scientific Support Team
https://www.abcam.com/index.html?pageconfig=technical&utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Body

[CCE4242373]


















Discover more at http://abcam.com/

Read More
Answer

Thank you for taking the time to reply with the information.

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 ab4193 is tested and covered by our 6 month guarantee for use in IHC-P 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.

Although the other antibody has worked well using this procedure, individual antibodies will often require optimization. Before deciding how to proceed, I would like to investigate this particular case further for you, and also obtain some further information for our quality records.

In order to do this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I have completed the sections for which information has already been provided.

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.

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



Order Details
Antibody code:

Lot number:


Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, non-specific staining etc.)

Sample (Species/Tissue/Cell Type/Cell Line etc.)

breast carcinoma tissue, skin (keratinocytes)

Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)


Antigen retrieval (Enzymatic method, Heat mediated technique etc.)

pH 6,0, antigen retrieval in PT-Link (DAKO)

What different incubation times and buffers been tried for antigen retreival to optimize?


Permeabilization step


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


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


1:500 (over night, 60 minutes and 1 hour of incubation)

Please confirm what other dilutions have been tried.

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


Is the current vial of secondary antibody working well with other primary antibodies?

Detection method


Positive and negative controls used (please specify)


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



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

Do you obtain the same results every time?
Yes No


What steps have you altered?


Additional Notes


We would appreciate if you are also able to provide and image which woudl help us to assess the results

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 these antibody.

I would like to reassure you that our antibodies are tested and covered by our 6 month guarantee for the tested species and applications listed on the individual datasheets. 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 would appreciate if you could let me know which application you have been using. Also which type of samples you have tested. Once I know which application you are using, I can obtain the protocol from the laboratory and will be pleased to send this on to you.

I would also appreciate if you are able to provide an image which would help us to assess the results.

Thank you for your time and cooperation. I look forward to hearing from you with the requested information.

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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