Recombinant Anti-Frizzled 4 antibody [22HCLC] (ab277797)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit recombinant multiclonal [22HCLC] to Frizzled 4
- Suitable for: ICC, IHC-P, WB
- Reacts with: Human
Overview
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Product name
Anti-Frizzled 4 antibody [22HCLC] -
Description
Rabbit recombinant multiclonal [22HCLC] to Frizzled 4 -
Host species
Rabbit -
Tested applications
Suitable for: ICC, IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment corresponding to Human Frizzled 4 aa 1-250.
Database link: Q9ULV1 -
Positive control
- WB: HCT 116, membrane extract of HCT116, T47D, membrane extract of T47D. IHC-P: Human kidney tissue. ICC:T-47D cells.
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General notes
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains.
Recombinant multiclonal antibodies offer the sensitivity of polyclonal antibodies by recognising multiple epitopes, along with consistency of a recombinant antibody.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.09% Sodium azide
Constituent: 99.91% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Recombinant Multiclonal -
Clone number
22HCLC -
Isotype
IgG -
Research areas
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab277797 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 |
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ICC |
Use a concentration of 2 µg/ml.
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IHC-P |
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 60 kDa.
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Notes |
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ICC
Use a concentration of 2 µg/ml. |
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 60 kDa. |
Target
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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
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Database links
- Entrez Gene: 8322 Human
- Omim: 604579 Human
- SwissProt: Q9ULV1 Human
- Unigene: 591968 Human
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Alternative names
- CD 344 antibody
- CD 344 antigen antibody
- CD344 antibody
see all
Images
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All lanes : Anti-Frizzled 4 antibody [22HCLC] (ab277797) at 2 µg/ml
Lane 1 : HCT 116 cell extract
Lane 2 : HCT116 membrane extract
Lane 3 : T47D cell extract
Lane 4 : T47D membrane extract
Secondary
All lanes : Goat anti-Rabbit IgG (H+L) Superclonal HRP conjugate at 1/2500 dilution
Predicted band size: 60 kDa -
Immunohistochemistry analysis of FZD4 showing staining in the cytoplasm of paraffin-embedded human kidney tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 minutes. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with a FZD4 Recombinant Rabbit Multiclonal Antibody (ab277797) diluted in 3% BSA-PBS at a dilution of 1/100 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
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Immunofluorescence was performed on fixed and permeabilized T-47D cells for detection of FZD-4 using Anti-FZD-4 Recombinant Rabbit Multiclonal Antibody (ab277797, 2 μg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (1/2000). Panel a) shows representative cells that were stained for detection and localization of FZD-4 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI. Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (1/300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of FZD-4. Panel e) represents control cells with no primary antibody to assess background.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (0)
ab277797 has not yet been referenced specifically in any publications.