The immunogen sequence is found in both AP2A2 and AP2A1. This antibody recognises a band of just under 100kD in multiple human cell lines (see picture), the band can be blocked with the immunising peptide.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500. Detects a band of approximately 100 kDa (predicted molecular weight: 104 , 108 kDa).
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 surrrounded 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 alpha subunit binds polyphosphoinositide-containing lipids, positioning AP-2 on the membrane. The AP-2 alpha subunit acts via its C-terminal appendage domain as a scaffolding platform for endocytic accessory proteins. The AP-2 alpha and AP-2 sigma subunits are thought to contribute to the recognition of the [ED]-X-X-X-L-[LI] motif.
Isoform A expressed in forebrain, skeletal muscle, spinal cord, cerebellum, salivary gland, heart and colon. Isoform B is widely expressed in tissues and also in breast cancer and in prostate carcinoma cells.
Belongs to the adaptor complexes large subunit family.
Cell membrane. Membrane > coated pit. AP-2 appears to be excluded from internalizing CCVs and to disengage from sites of endocytosis seconds before internalization of the nascent CCV.
Plasma membrane adaptor HA 2/AP 2 adaptin alpha A subunit antibody
Plasma membrane adaptor HA2/AP2 adaptin alpha A subunit antibody
- AP2 alpha antibody (ab3707)
Western blot using ab3707 at 1/500.
Lane 1: HeLa Nuclear Extract Lane 2: HeLa Whole Cell Lysate Lane 3: 293 Whole Cell Lysate Lane 4: A431 Whole Cell Lysate Lane 5: Jurkat Whole Cell Lysate Lane 6: HeLa Nuclear Extract + blocking/immunising peptide Lane 7: HeLa Whole Cell Lysate + blocking/immunising peptide Lane 8: 293 Whole Cell Lysate + blocking/immunising peptide Lane 9: A431 Whole Cell Lysate + blocking/immunising peptide Lane 10: Jurkat Whole Cell Lysate + blocking/immunising peptide
AP2 alpha proteins (AP2A2 and AP2A1) have predicted molecular weights of 104 and 107kD. The band at just below 97kD (that is blocked by the immunising peptide) represents AP2 alpha, the lower bands are not blocked completely by the immunising peptide and are believed to be non-specific.
Secondary ab: Rabbit polyclonal to Goat IgG HRP ab6741 (1/5000) Exposure time: 1 minute. Lysates at 20µg/lane. Blocking peptide at 1µg/ml.