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A4325 Sigma

Monoclonal Anti-α-Adaptin antibody produced in mouse

clone 100/2, ascites fluid

Synonym: Anti-AP-2

Properties

Related Categories AD-AD, Alphabetical Index, Antibodies, Primary Antibodies
species reactivity   bovine, human, rat
application(s)   western blot: 1:200 using a crude preparation of synaptic vesicles obtained from rat cerebral cortex
clone   100/2, monoclonal
antibody form   ascites fluid
isotype   IgG2a
contains   15 mM sodium azide
shipped in   dry ice
storage temp.   −20°C
Gene Information   human ... AP2A1(160)
rat ... Ap2a1(308578)
biological source   mouse
conjugate   unconjugated

Description

Immunogen

AP-2 adaptor polypeptides from bovine brain.

General description

Clathrin-coated vesicles facilitate the selective internalization of membrane receptors for lysosomal enzymes from the trans-Golgi network to a pre-lysosomal compartment. Besides clathrin, coated vesicle populations contain the adaptor complexes AP-1 and AP-2, also known as HA-I (HA1) adaptor and HA-II (HA2) adaptor or Assemble Protein 1 (AP1) and Assemble Protein 2 (AP2), respectively. These adaptor proteins mediate the interaction between clathrin and membrane components.

The antibody reacts with the 105 and 110 kDa α-subunits of adaptor complex AP-2 in bovine and rat brain preparations. It reacts with polypeptides of ~100 kDa in bovine liver, human heart fibroblasts, and Madin-Darby bovine kidney cultured cells (MDBK), but not with any components in the 110-115 kDa range from these sources or from rat pheochromocytoma cultured cells (PC12), neuroblastoma, or astrocytes. This suggests that the 110 and 115 kDa polypeptides may be specific variants that occur only in some cell types of brain. The antibody stains the intact α-adaptor, as well as the 37 and 40 kDa fragments, but not the 63-66 kDa group of α-fragments obtained by trypsin cleavage of the α-subunit molecules.

Physical form

The product is provided as ascites fluid with 0.1% sodium azide as a preservative.

Application

Immunofluorescence of MDCK cells fixed in methanol was performed using anti-AP1 at a dilution of 1:250 and also western blot at a dilution of 1:1000.

Mouse monoclonal anti-α-adaptin antibody has been used as a positive control in immunoblotting assays. It has also been used as a primary antibody to detect AP2 levels in tetracycline-repressible stable cell lines.

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