The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 2.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 0.1 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 63.6 kDa). Preliminary experiments gave an approx. 23kDa band in Human Colon, Heart and Ovary lysates after 0.1µg/ml antibody staining. Please note that currently we cannot find an explanation in the literature for the band we observe given the calculated size of 63.6kDa according to NP_001457.1. The 23kDa band was successfully blocked by incubation with the immunizing peptide.
Use a concentration of 10 µg/ml.
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.
Widely expressed. In the adult, mainly found in heart, placenta, skeletal muscle, lung, kidney, pancreas, prostate, testis, ovary and colon. In the fetus, expressed in brain, lung and kidney. Low levels in fetal liver.
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 ab109094 stained A549 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA, 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells. The cells were then incubated with the antibody ab109094 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 donkey anti- goat (ab96931) 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.