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Anti-AAK1 Antibody

Rabbit polyclonal antibody to AAK1

Western blot analysis of AAK1 expression in Hela (A) whole cell lysates.
Immunohistochemical analysis of AAK1 staining in human brain formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Immunofluorescent analysis of AAK1 staining in HepG2 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a hidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IF/IC, IHC
Primary Accession	Q2M2I8
Other Accession	Q3UHJ0
Reactivity	Human, Mouse, Rat, Bovine
Host	Rabbit
Clonality	Polyclonal
Calculated MW	103885 Da

Additional Information
Gene ID	22848
Other Names	KIAA1048; AP2-associated protein kinase 1; Adaptor-associated kinase 1
Target/Specificity	KLH-conjugated synthetic peptide encompassing a sequence within the center region of human AAK1. The exact sequence is proprietary.
Dilution	WB~~WB (1/500 - 1/2000), IHC (1/50 - 1/200), IF/IC (1/50 - 1/100) IF/IC~~N/A IHC~~WB (1/500 - 1/2000), IHC (1/50 - 1/200), IF/IC (1/50 - 1/100)
Format	Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.09% (W/V) sodium azide.
Storage	Store at -20 °C.Stable for 12 months from date of receipt

Protein Information
Name	AAK1
Synonyms	KIAA1048
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 (PubMed:11877457, PubMed:11877461, PubMed:12952931, PubMed:14617351, PubMed:17494869, PubMed:25653444). Isoform 1 and isoform 2 display similar levels of kinase activity towards AP2M1 (PubMed:17494869). Preferentially, may phosphorylate substrates on threonine residues (PubMed:11877457, PubMed:18657069). Regulates phosphorylation of other AP-2 subunits as well as AP-2 localization and AP-2-mediated internalization of ligand complexes (PubMed:12952931). Phosphorylates NUMB and regulates its cellular localization, promoting NUMB localization to endosomes (PubMed:18657069). Binds to and stabilizes the activated form of NOTCH1, increases its localization in endosomes and regulates its transcriptional activity (PubMed:21464124).
Cellular Location	Cell membrane {ECO:0000250\|UniProtKB:F1MH24}; Peripheral membrane protein {ECO:0000250\|UniProtKB:F1MH24}. Membrane, clathrin-coated pit. Presynapse {ECO:0000250\|UniProtKB:P0C1X8}. 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. {ECO:0000250\|UniProtKB:P0C1X8}
Tissue Location	Detected in brain, heart and liver. Isoform 1 is the predominant isoform in brain.