Overview

  • Product name

  • Description

    Goat polyclonal to Frizzled 4
  • Host species

    Goat
  • Specificity

    Tested in human and expected to react in rat given sequence similarity
  • Tested applications

    Suitable for: WB, ELISAmore details
  • Species reactivity

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

    Synthetic peptide:

    DEEERRCDPIRIS

    , corresponding to amino acids 39-51 of Human Frizzled 4 (NP_036325.2).

  • Positive control

    • Approx 60kDA band observed in Human Colon and Small Intestine

Properties

Applications

Our Abpromise guarantee covers the use of ab77724 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 0.3 - 1 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).
ELISA 1/16000.

Target

  • Function

    Receptor for Wnt proteins. Most of frizzled receptors are coupled to the beta-catenin (CTNNB1) canonical signaling pathway, which leads to the activation of disheveled proteins, inhibition of GSK-3 kinase, nuclear accumulation of beta-catenin (CTNNB1) and activation of Wnt target genes. Plays a critical role in retinal vascularization by acting as a receptor for Wnt proteins and norrin (NDP). In retina, it can be both activated by Wnt protein-binding, but also by a Wnt-independent signaling via binding of norrin (NDP), promoting in both cases beta-catenin (CTNNB1) accumulation and stimulation of LEF/TCF-mediated transcriptional programs. A second signaling pathway involving PKC and calcium fluxes has been seen for some family members, but it is not yet clear if it represents a distinct pathway or if it can be integrated in the canonical pathway, as PKC seems to be required for Wnt-mediated inactivation of GSK-3 kinase. Both pathways seem to involve interactions with G-proteins. May be involved in transduction and intercellular transmission of polarity information during tissue morphogenesis and/or in differentiated tissues.
  • Tissue specificity

    Almost ubiquitous. Largely expressed in adult heart, skeletal muscle, ovary, and fetal kidney. Moderate amounts in adult liver, kidney, pancreas, spleen, and fetal lung, and small amounts in placenta, adult lung, prostate, testis, colon, fetal brain and liver.
  • Involvement in disease

    Defects in FZD4 are the cause of vitreoretinopathy exudative type 1 (EVR1) [MIM:133780]; also known as autosomal dominant familial exudative vitreoretinopathy (FEVR) or Criswick-Schepens syndrome. EVR1 is a disorder of the retinal vasculature characterized by an abrupt cessation of growth of peripheral capillaries, leading to an avascular peripheral retina. This may lead to compensatory retinal neovascularization, which is thought to be induced by hypoxia from the initial avascular insult. New vessels are prone to leakage and rupture causing exudates and bleeding, followed by scarring, retinal detachment and blindness. Clinical features can be highly variable, even within the same family. Patients with mild forms of the disease are asymptomatic, and their only disease-related abnormality is an arc of avascular retina in the extreme temporal periphery.
  • Sequence similarities

    Belongs to the G-protein coupled receptor Fz/Smo family.
    Contains 1 FZ (frizzled) domain.
  • Domain

    Lys-Thr-X-X-X-Trp motif interacts with the PDZ doman of Dvl (Disheveled) family members and is involved in the activation of the Wnt/beta-catenin signaling pathway.
    The FZ domain is involved in binding with Wnt ligands.
  • Cellular localization

    Membrane. Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • CD 344 antibody
    • CD 344 antigen antibody
    • CD344 antibody
    • CD344 antigen antibody
    • EVR1 antibody
    • FEVR antibody
    • Frizzled (Drosophila) homolog 4 antibody
    • Frizzled 4 antibody
    • Frizzled family receptor 4 antibody
    • Frizzled homolog 4 (Drosophila) antibody
    • Frizzled-4 antibody
    • Fz 4 antibody
    • Fz-4 antibody
    • Fz4 antibody
    • FZD4 antibody
    • FZD4_HUMAN antibody
    • FZD4S antibody
    • FzE4 antibody
    • GPCR antibody
    • hFz4 antibody
    • MGC34390 antibody
    • WNT receptor frizzled 4 antibody
    see all

Images

  • Anti-Frizzled 4 antibody (ab77724) at 1 µg/ml + Small Intestine (Ileum) cell lysate at 35 µg

    Predicted band size: 60 kDa
    Observed band size: 60 kDa

References

This product has been referenced in:

  • Formosa A  et al. MicroRNAs, miR-154, miR-299-5p, miR-376a, miR-376c, miR-377, miR-381, miR-487b, miR-485-3p, miR-495 and miR-654-3p, mapped to the 14q32.31 locus, regulate proliferation, apoptosis, migration and invasion in metastatic prostate cancer cells. Oncogene N/A:N/A (2013). Human . Read more (PubMed: 24166498) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question

Please see the answers to your questions from the lab below:
- Have you tried whole cells lysates with this antibody? How were the results? I am assuming the problem might be with membrane fractions rather than antibody.
YES. WE SENT YOU THE IMAGES OF WHOLE CELL LYSATES. THAT WAS OUR FIRST SAMPLE TRIAL. WE MOVED ONTO MEMBRANE FRACTIONS TO CLEAR UP SOME BACKGROUND. ALTHOUGH IT SEEMED TO HELP, WE CANNOT ACHIEVE OUR GOAL OF MAKING A CLEAN IMAGE FOR PUBLICATION
>- Have you tried higher dilution of antibody? WITH HIGHER ANTIBODY
>DILUTION,
THE BAND OF INTEREST IS MORE FADED, WHICH REQUIRES A LONGER EXPOSURE, AND THUS INCREASES BACKGROUND NONETHELESS.
>
>I have following further protocol optimization suggestions;
>
>- Block membrane with 5% BSA, for 2 hours. WE TRIED THIS TODAY. IT
>WORSENED
THE PROBLEM.
>- Try more diluted primary antibody. Probably 1/1000 dilution for 4 hours.
Increasing the dilution sometime solves the background problem. WE BEGAN INITIALLY WITH HIGHER DILUTIONS.
>- CD344 is a multi pass membrane protein. Please do not boil the
>samples
instead heat samples at 60-70C for 15 minutes. HAVE TRIED THIS TODAY. NO CHANGE
>- Try using loading control. WE HAVE A CONTROL. PC3 CELLS TREATED WITH
>SIRNA
FOR FZD4 (DHARMACON). WE HAVE A STRONG PHENOTYPE WITH SILENCING, THEREFORE THE PROTEIN IS EXPRESSED.
>- We have tested this antibody using MCF7 and fetal liver lysates. Try
>these
cell lines as positive control. SEE PREVIOUS QUESTION
The lab are anxious to know if/when we should receive as replacement the item ab77724?
Regards,

Read More
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1137090.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

Read More

Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.
I have read the details you have kindly provided and have following further questions for better understanding of the problem;
- Have you tried whole cells lysates with this antibody? How were the results? I am assuming the problem might be with membrane fractions rather than antibody.
- Have you tried higher dilution of antibody?
I have following further protocol optimization suggestions;
- Block membrane with 5% BSA, for 2 hours.
- Try more diluted primary antibody. Probably 1/1000 dilution for 4 hours. Increasing the dilution sometime solves the background problem.
- CD344 is a multi pass membrane protein. Please do not boil the samples instead heat samples at 60-70C for 15 minutes.
- Try using loading control.
- We have tested this antibody using MCF7 and fetal liver lysates. Try these cell lines as positive control.
Finally, could you provide the purchase order number?
I am sure these suggestions would be helpful. Should you have any question or the results do not improve, please do not hesitate to contact me again.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up