The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 10 µg/ml.
1/2000 - 1/5000. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).
Regulates Wnt proteins sorting and secretion in a feedback regulatory mechanism. This reciprocal interaction plays a key role in the regulation of expression, subcellular location, binding and organelle-specific association of Wnt proteins. Plays also an important role in establishment of the anterior-posterior body axis formation during development.
ICC/IF image of ab72385 stained SKNSH cells. The cells were 4% formaldehyde 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 ab72385 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- chicken (ab96947) IgY (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.
Western blot - GPR177 antibody (ab72385)
All lanes : Anti-GPR177 antibody (ab72385) at 1/2000 dilution
Lane 1 : Molecular weight markers Lane 2 : HEK293 cell lysate transfected with GPR177-GFP
Predicted band size: 62 kDa Observed band size: 62 kDa
The HEK293 cell protein lysate contains both transfected GPR177-GFP fusion protein (75 kDa) and endogenous GPR177 protein (~62 kDa).
de Groot RE et al. Retromer dependent recycling of the wnt secretion factor wls is dispensable for stem cell maintenance in the Mammalian intestinal epithelium. PLoS One8:e76971 (2013).
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