Overview

  • Product name

  • Description

    Rabbit polyclonal to Frizzled 4
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Horse, Dog, Human
    Predicted to work with: Chicken, Guinea pig, Cow, Cat
  • Immunogen

    Synthetic peptide corresponding to Human Frizzled 4 aa 488-537 (internal sequence).
    Sequence:

    GITSGMWIWSAKTLHTWQKCSNRLVNSGKVKREKRGNGWVKPGKGSETVV


    Database link: NP_036325

  • Positive control

    • Fetal liver lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab83042 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 Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 4 - 8 µg/ml.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 60 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

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 (ab83042) at 1 µg/ml + fetal liver lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 60 kDa
    Observed band size: 65 kDa
    why is the actual band size different from the predicted?



    Gel concentration 12%
  • ab83042 staining Frizzled 4 in rat brain tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/10000 in TBS/BSA/azide) for 2 hours at 21°C. A Biotin-conjugated goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab83042 stained MCF7 cells. The cells were 4% PFA fixed (10mins) 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 (ab83042, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

  • Immunohistochemistry (Formalin/PFa-fixed paraffin-embedded sections) analysis of human skin tissue labelling Frizzeled 4 with ab83042 at 4-8µg/ml.

  • ab83042 staining Frizzled 4 in human epithelium tissue sections (top image - skin, bottom image - bronchiolar epithelium) by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/6000 in TBS/BSA/azide) for 2 hours at 21°C. A Biotin-conjugated goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:

  • Chen L  et al. MicroRNA-101 inhibits cell migration and invasion in bladder cancer via targeting FZD4. Exp Ther Med 17:1476-1485 (2019). Read more (PubMed: 30680031) »
  • Ota H  et al. Identification of the X-linked germ cell specific miRNAs (XmiRs) and their functions. PLoS One 14:e0211739 (2019). Read more (PubMed: 30707741) »
See all 11 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Monkey Cell lysate - whole cell (COS7)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
COS7
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Nov 07 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted May 25 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted May 25 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted May 24 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Human Tissue sections (Epithelium)
Specification
Epithelium
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Feb 19 2015

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

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