The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use at an assay dependent concentration. PubMed: 23251385
Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 64 kDa).
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Receptor for Wnt proteins. Most of frizzled receptors are coupled to the beta-catenin canonical signaling pathway, which leads to the activation of disheveled proteins, inhibition of GSK-3 kinase, nuclear accumulation of beta-catenin and activation of Wnt target genes. 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.
High expression in adult skeletal muscle and fetal kidney, followed by fetal lung, adult heart, brain, and placenta. Specifically expressed in squamous cell esophageal carcinomas.
Belongs to the G-protein coupled receptor Fz/Smo family. Contains 1 FZ (frizzled) domain.
Lys-Thr-X-X-X-Trp motif is involved in the activation of the Wnt/beta-catenin signaling pathway. The FZ domain is involved in binding with Wnt ligands.
ICC/IF image of ab64636 stained MCF7 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 (ab64636, 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 5µg/ml.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Frizzled 7 antibody (ab64636)This image is courtesy of an abreview submitted by Antibody Solutions Ltd.
ab64636 (1/250) staining Frizzled 7 in paraffin-embedded Human kidney (papillary cancer) tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. The secondary antibody was goat anti rabbit conjugated to HRP. For further experimental details please refer to abreview.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Frizzled 7 antibody (ab64636)Image from Gonzalez P et al., PLoS One. 2012;7(12):e50793. Fig 5.; doi: 10.1371/journal.pone.0050793. Epub 2012 Dec 10
Immunohistochemical image of ab64636 (dilution 1/2000) staining in rat spinal cord PFA fixed frozen tissue section, performed on an Olympus BX61 Motorized Research Microscope. The scale bar represents 50 µm.
Western blot - Anti-Frizzled 7 antibody (ab64636)
All lanes : Anti-Frizzled 7 antibody (ab64636) at 1 µg/ml
Lane 1 : Skeletal Muscle (Mouse) Tissue Lysate Lane 2 : Spinal Cord (Rat) Tissue Lysate Lane 3 : Human brain tissue lysate - total protein (ab29466) Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 64 kDa Additional bands at: 90 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 10 minutes
Frizzled 7 has a predicted molecular weight of 64 kDa, however as it contains a number of potential glycosylation sites (SwissProt data) it is thought that this is the reason it is running at 90kDa.
Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
IHC image of Frizzled 7 staining in human skeletal muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab64636, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Cao TT et al. FZD7 is a novel prognostic marker and promotes tumor metastasis via WNT and EMT signaling pathways in esophageal squamous cell carcinoma. Oncotarget8:65957-65968 (2017).
Read more (PubMed: 29029485) »
Proto MC et al. Inhibition of Wnt/ß-Catenin pathway and Histone acetyltransferase activity by Rimonabant: a therapeutic target for colon cancer. Sci Rep7:11678 (2017).
Read more (PubMed: 28916833) »