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 a concentration of 1 µg/ml. Detects a band of approximately 168 kDa (predicted molecular weight: 93 kDa).
RelevanceAP180 is a clathrin binding phospho-protein and faciliates the formation of clathrin coats. This protein is also known as pp155, NP185, F1-20, and AP-3. During neurotransmitter release synaptic vesicles fuse with the presynaptic plasma membrane. A whole protein machinery consisting of amphiphysin, clathrin, endophilin and synaptojanin is involved in the subsequent endocytotic recycling of the synaptic vesicles.
Cellular localizationComponent of the coat surrounding the cytoplasmic face of coated vesicles in the plasma membrane.
AP180 is a brain specific protein that contains a number of potential phosphorylation sites (SwissProt data) which may explain it running at a higher molecular weight than predicted. AP180 has also been shown to run in western blot at approximately 180 kDa in numerous publications, e.g. Am J Physiol Cell Physiol 285: C995-C1008, 2003 (PMID: 14532018).
ICC/IF image of ab33898 stained PC12 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 (ab33898, 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).
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-AP180 antibody (ab33898)This image is courtesy of an abreview submitted by Sophie Pezet
Immunostaining of Mouse Striatum suing Ab33898. The tissues are from perfused fixed animals perfused with 4% PFA and later postfixed overnight in the same fixative. They were cryoprotected in 30% sucrose and cut using a cryostat and stained by direct fluorescence.