Polyclonal antibody to Phospho-AP2M1-T156
Human, Mouse, Rat
AP2M1 antibody; AP50 antibody; CLAPM1 antibody; mu2 antibody; AP-2 complex subunit mu antibody
Antigen: A synthetic phosphorylated peptide around T156 of human AP2M1 (NP_001020376.1).
PBS with 0.02% sodium azide, 50% glycerol, pH7.3.
Store at -20℃. Avoid freeze / thaw cycles.
WB 1:500 - 1:2000
Western blot - Phospho-AP2M1-T156 pAb
Western blot analysis of extracts of 293T and NIH/3T3 cells, using Phospho-AP2M1-T156 antibody at 1:1000 dilution. 293T cells were treated by Calyculin A for 30 minutes after serum-starvation overnight.NIH/3T3 cells were treated by Hydrogen Peroxide (2nM) for 15 minutes after serum-starvation overnight.Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25ug per lane.Blocking buffer: 3% BSA.Detection: ECL Basic Kit .Exposure time: 30s.
Component of the adaptor protein complex 2 (AP-2). Adaptor protein complexes function in protein transport via transport vesicles in different membrane traffic pathways. Adaptor protein complexes are vesicle coat components and appear to be involved in cargo selection and vesicle formation. AP-2 is involved in clathrin-dependent endocytosis in which cargo proteins are incorporated into vesicles surrounded by clathrin (clathrin-coated vesicles, CCVs) which are destined for fusion with the early endosome. The clathrin lattice serves as a mechanical scaffold but is itself unable to bind directly to membrane components. Clathrin-associated adaptor protein (AP) complexes which can bind directly to both the clathrin lattice and to the lipid and protein components of membranes are considered to be the major clathrin adaptors contributing the CCV formation. AP-2 also serves as a cargo receptor to selectively sort the membrane proteins involved in receptor-mediated endocytosis. AP-2 seems to play a role in the recycling of synaptic vesicle membranes from the presynaptic surface. AP-2 recognizes Y-X-X-[FILMV] (Y-X-X-Phi) and [ED]-X-X-X-L-[LI] endocytosis signal motifs within the cytosolic tails of transmembrane cargo molecules. AP-2 may also play a role in maintaining normal post-endocytic trafficking through the ARF6-regulated, non-clathrin pathway. The AP-2 mu subunit binds to transmembrane cargo proteins; it recognizes the Y-X-X-Phi motifs. The surface region interacting with to the Y-X-X-Phi motif is inaccessible in cytosolic AP-2, but becomes accessible through a conformational change following phosphorylation of AP-2 mu subunit at 'Tyr-156' in membrane-associated AP-2. The membrane-specific phosphorylation event appears to involve assembled clathrin which activates the AP-2 mu kinase AAK1 (By similarity). Plays a role in endocytosis of frizzled family members upon Wnt signaling (By similarity).
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