Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab4193 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
ELISA 1/1000.
ICC/IF Use a concentration of 5 µg/ml.
IHC-P 1/800. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Boil the sections for 20-30 minutes in water bath. We also recomend trying different antigen retrieval methods with different buffers if one does not work.

WB 1/230. Predicted molecular weight: 37 kDa.

Target

  • Function
    Soluble frizzled-related proteins (sFRPS) function as modulators of Wnt signaling through direct interaction with Wnts. They have a role in regulating cell growth and differentiation in specific cell types. SFRP1 decreases intracellular beta-catenin levels (By similarity). Has antiproliferative effects on vascular cells, in vitro and in vivo, and can induce, in vivo, an angiogenic response. In vascular cell cycle, delays the G1 phase and entry into the S phase (By similarity). In kidney development, inhibits tubule formation and bud growth in metanephroi (By similarity). Inhibits WNT1/WNT4-mediated TCF-dependent transcription.
  • Tissue specificity
    Widely expressed. Absent from lung, liver and peripheral blood leukocytes. Highest levels in heart and fetal kidney. Also expressed in testis, ovary, fetal brain and lung, leiomyomal cells, myometrial cells and vascular smooth muscle cells. Expressed in foreskin fibroblasts and in keratinocytes.
  • Sequence similarities
    Belongs to the secreted frizzled-related protein (sFRP) family.
    Contains 1 FZ (frizzled) domain.
    Contains 1 NTR domain.
  • Domain
    The FZ domain is involved in binding with Wnt ligands.
  • Cellular localization
    Secreted. Cell membrane or extracellular matrix-associated. Released by heparin-binding.
  • Information by UniProt
  • Database links
  • Alternative names
    • Frizzled related protein 1 antibody
    • FRP 1 antibody
    • FRP antibody
    • FRP-1 antibody
    • FRP1 antibody
    • FrzA antibody
    • SARP 2 antibody
    • SARP-2 antibody
    • SARP2 antibody
    • Secreted apoptosis related protein 2 antibody
    • Secreted apoptosis-related protein 2 antibody
    • Secreted frizzled related protein 1 antibody
    • Secreted frizzled related protein antibody
    • Secreted frizzled-related protein 1 antibody
    • SFRP 1 antibody
    • sFRP-1 antibody
    • SFRP1 antibody
    • SFRP1_HUMAN antibody
    see all

Images

  • ab4193 staining human dermal hypertrophic scar tissue sections by IHC-P.  Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval prior to blocking (with a commercial protein block) and incubating with the primary antibody (diluted 1/800) for 20 minutes.  A HRP-conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

  • Western blot using ab4193 at 1/230 (o/n 4°C) on lysates from cultured small airway epithelial cells.

    Expected band size: 37kD.

    Western blot using ab4193 at 1/230 (o/n 4°C) on lysates from cultured small airway epithelial cells.

  • ICC/IF image of ab4193 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab4193, 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.

  • ab4193 staining SFRP1 by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) in human breast cancer cells, injected subcutaneously into mice and grown until the tumour was a palpable size. Tissue underwent paraformaldehyde fixation before heat mediated antigen retrieval in 10mM sodium citrate- EDTA (pH 6). The primary antibody was diluted 1/1000 and incubated with sample for 17 hours at 4°C in 10% Casein. A Biotin conjugated goat polyclonal to rabbit IgG was used at dilution at 1/200 as secondary antibody.

    See Abreview

  • ab4193 detecting SFRP1 in Human Tear samples (purified protein) by sandwich ELISA. Protein was captured and Samples were blocked with 3% milk for 2 hours at 25°C and incubated with ab4193 (1/1000 in PBS ) for 1 hour at 37°C. An alkaline phosphatase Goat anti-rabbit IgG polyclonal (1/15000) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Hawkshaw NJ  et al. Identifying novel strategies for treating human hair loss disorders: Cyclosporine A suppresses the Wnt inhibitor, SFRP1, in the dermal papilla of human scalp hair follicles. PLoS Biol 16:e2003705 (2018). Read more (PubMed: 29738529) »
  • Chen Y  et al. Obesity-associated miR-27a upregulation promotes hepatocellular carcinoma metastasis through suppressing SFRP1. Onco Targets Ther 11:3281-3292 (2018). Read more (PubMed: 29910623) »
See all 27 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Answer

Your protocol appears to be correct. The only IHC data we have is from customers who used citrate heat-mediated retrieval and dilutions of 1/1000 or 1/800. The samples were hypertrophic scar and a subcutaneous implant of MDA-MB 436 breast cancer cells. Are you confident that your secondary antibody is effective? It should be anti-rabbit IgG. The SFRP1 protein is located in the plasma membrane but it is also secreted, which may account for staining in the extra-cellular matrix.

Here are links to the reviews:

https://www.abcam.com/SFRP1-antibody-ab4193-reviews.html?intabreviewid=15717

https://www.abcam.com/SFRP1-antibody-ab4193-reviews.html?intabreviewid=9877

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Answer

The highest expression is in the heart and kidney. For an example of a strong stain of heart muscle tissue, please see the image on the datasheet for a different SFRP1 antibody, ab126613:

Click here (or use the following: https://www.abcam.com/index.html?datasheet=126613).

For ab4193, we recommend a heat-mediated antigen retrieval, based on the customer review from 2008:

https://www.abcam.com/SFRP1-antibody-ab4193-reviews.html?intabreviewid=9877

The reviewer used a DAKO retrieval solution but a 10mM sodium citrate buffer, pH 6.0, should be sufficient.

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Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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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.

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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/

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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

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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.

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Answer

Este es un mail de confirmación del nuevo pedido, consistente en el reemplazo completamente gratuito, del anticuerpo ab4193 con número de pedido XXX.

Os llegará un mail de confirmación del nuevo pedido, si necesitáis información sobre el estado del mismo no dudéis en contactarnos haciendo referencia al número indicado.

Lamento mucho las molestias ocasionadas. No dudéis en contactarnos para cualquier otra duda o información.

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Answer

Gracias por contactarnos y alertarnos del problema sufrido con este anticuerpo.

Efectivamente los resultados obtenidos con este nuevo lote no se corresponden con los esperados, y le pido disculpas por las molestias que esto haya podido ocasionar.

Dado que nuestros productos tienen 6 meses de garantía en caso de no funcionar como se especifica en su datasheet, os reemplazaré el producto por un lote diferente. Sin embargo, no tenemos actualmente en stock el lote que os dió buenos resultados en el pasado.

Tenemos otros que difieren de los dos ya testados, si me confirmáis que queréis recibir el reemplazo por uno de ellos, crearé el pedido de inmediato. Alternativamente podemos sustituir el anticuerpo por otro diferente de nuestro catálogo, o reembolsaros el importe.

Quedo a la espera de recibir su respuesta para proceder.

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Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Human Purified protein (Tears)
Specification
Tears
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Type
Sandwich (Detection)

Abcam user community

Verified customer

Submitted Aug 24 2012

Answer

Thank you for your email.

I apologize for the delay as it took a while to get back to you. Unfortunately the publication team at Abcam is unable to trace back the antibodies catalogue number the author have used. They can only trace one abID which is ab4193.

I would suggest contacting the author for this information. I am sure they will be able to release this information.

I hope this information is nevertheless helpful. Should you have any further question please do not hesitate to contact me.

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1-10 of 19 Abreviews or Q&A

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

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