|Application ||WB, IF, ICC, E|
|Other Accession||NP_055726, 148277037|
|Reactivity||Human, Mouse, Rat|
|Calculated MW||103885 Da|
|Application Notes||Aak1 antibody can be used for detection of Aak1 by Western blot at 1 - 2 µg/mL. Antibody can also be used for immunocytochemistry starting at 5 µg/mL. For immunofluorescence start at 20 µg/mL.|
|Reconstitution & Storage||Aak1 antibody can be stored at 4℃ for three months and -20℃, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.|
|Precautions||Aak1 Antibody is for research use only and not for use in diagnostic or therapeutic procedures.|
|Function||Regulates clathrin-mediated endocytosis by phosphorylating the AP2M1/mu2 subunit of the adaptor protein complex 2 (AP-2) which ensures high affinity binding of AP-2 to cargo membrane proteins during the initial stages of endocytosis. Isoform 1 and isoform 2 display similar levels of kinase activity towards AP2M1. Regulates phosphorylation of other AP-2 subunits as well as AP-2 localization and AP-2-mediated internalization of ligand complexes. Phosphorylates NUMB and regulates its cellular localization, promoting NUMB localization to endosomes. Binds to and stabilizes the activated form of NOTCH1, increases its localization in endosomes and regulates its transcriptional activity.|
|Cellular Location||Cell membrane; Peripheral membrane protein. Membrane, clathrin-coated pit. Note=Active when found in clathrin-coated pits at the plasma membrane. In neuronal cells, enriched at presynaptic terminals. In non-neuronal cells, enriched at leading edge of migrating cells (By similarity).|
|Tissue Location||Detected in brain, heart and liver. Isoform 1 is the predominant isoform in brain|
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Provided below are standard protocols that you may find useful for product applications.
Aak1 Antibody: AP2-associated protein kinase 1 (Aak1) is a member of the Ark1/Prk1 subfamily of Ser/Thr protein kinases that are thought to regulate endocytosis by phosphorylating the accessory endocytic components. Aak1 interacts with and phosphorylates the mu2 subunit of the AP-2 complex, which promotes binding of the AP-2 to tyrosine based (Yxxphi) internalization motif-containing receptors and subsequent receptor endocytosis. At least two isoforms of Aak1 are known to exist; the longer isoform contains an extended carboxy-terminus that contains an additional clathrin-binding domain. Overexpression of this long isoform or Aak1 depletion by RNA interference impairs transferrin recycling from the early/sorting endosome, suggesting that Aak1 functions at multiple steps of the endosomal pathway by regulating transferrin internalization and its recycling back to the plasma membrane.
Connor SD and Schmid SL. Identification of an adaptor-associated kinase, AAK1, as a regulator of clathrin-mediated endocytosis. J. Cell Biol.2002; 156:921-9.
Smythe E and Ayscough KR. The Ark1/Prk1 family of protein kinases. Regulators of endocytosis and the actin skeleton. EMBO Rep.2003; 4:246-51.
Ricotta D, Connor SD, Schmid SL, et al. Phosphorylation of the AP2 m2 subunit by AAK1 mediates high affinity binding to membrane protein sorting signals. J. Cell Biol.2002; 156:791-5.
Connor SD and Henderson DM. A novel AAK1 splice variant functions at multiple steps of the endocytic pathway. Mol. Biol. Cell2007; 18:2698-706.
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