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A4450 Sigma

Monoclonal Anti-β1 and β2-Adaptins antibody produced in mouse

clone 100/1, ascites fluid

Synonym: Anti-AP-1 and AP-2

Properties

Related Categories AD-AD, Alphabetical Index, Antibodies, Primary Antibodies
species reactivity   bovine, rat, human
application(s)   indirect immunofluorescence: suitable
  microarray: suitable
  western blot: 1:200 using rat brain extract
clone   100/1, monoclonal
antibody form   ascites fluid
isotype   IgG1
contains   15 mM sodium azide
shipped in   dry ice
storage temp.   −20°C
Gene Information   human ... AP1B1(162), AP2B1(163)
rat ... Ap1b1(29663), Ap2b1(140670)
biological source   mouse
conjugate   unconjugated

Description

Immunogen

AP-2 adaptor protein 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 in immunoblotting of bovine and rat brain preparations, with a major band ~115 kDa and a minor component about 110 kDa of the adaptor complex AP-1, and with the 104 or 106 and 112 kDa polypeptides of the adaptor complex AP-2. The antibody 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 occurring only in some cell types of brain. The antibody may be used for immunofluorescent staining of clathrin-coated membranes at the plasma membrane and in the Golgi region of methanol-fixed MDBK cells.

Physical form

The antibody 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-β1 and β2-adaptins can be used for western blot (1:200) assays using rat brain extract. The antibody is also suitable for use in indirect immunofluorescence and microarray analyses. Furthermore, monoclonal anti-β1/β2-adaptins have been used for double staining methods to detect clathrin-coated regions of the plasma membrane in rat brain cells.

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