Adaptin, alpha

Clathrin-mediated endocytosis is the pathway by which many receptors for nutrients and hormones are internalized to be recycled or down-regulated. During formation of clathrin coated membranes, clathrin co-assembles with heterotetrameric molecules known as assembly polypeptides (APs) or adaptors which form a layer of protein coat between the clathrin lattice and the membrane. There are two characterized adaptors AP1 and AP2. AP1 is associated with clathrin coated vesicles at the trans-Golgi network and AP2 is associated with the endocytic clathrin coated vesicles at the plasma membrane and has been shown to specifically interact with Shc and EGF receptor. AP2 is composed of four subunits, two separate 100kD gene products with similar domain structures (alpha and beta adaptin) and a 50 and 17kD subunit. There are two alpha-adaptin genes, alpha A and alpha C which have a tissue specific pattern of expression. \n\nApplications: \nSuitable for use in Immunoprecipitation, Blocking, Immunofluorescence and Western Blot. Other applications not tested.\n\nRecommended Dilutions: \nImmunofluorescence: 1:200\nWestern Blot: 1:1000. Detects an ~100kD doublet representing the two isoforms of alpha adaptin in rat brain, though alpha C is the dominant form seen.\nBlocking: Microinjection of cells with A0859-90D results in variable blockade of endocytosis. \nOptimal dilutions to be determined by researher.\n\nStorage and Stability: \nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

A0859-90D

Size

100ul

Applications

IF, IP, WB

Hosts

Mouse

Reactivities

Hum, Mouse, Rat, Bov

Form

Supplied as a liquid in PBS, 0.05% sodium azide.

P Type

Mab

Purity

IgG fraction

Isotype

IgG1

References

1. Chin, D.J., et al.: 100-kDa polypeptides in peripheral clathrin-coated vesicles are required for receptor-mediated endocytosis. PNAS USA 86: 9289-93 (1989). 2. Chang, M.P., et al.: Adaptor self-aggregation, adaptor-receptor recognition and binding of alpha-adaptin subunits to the plasma membrane contribute to recruitment of adaptor (AP2) components of clathrin-coated pits. EMBO J. 12: 2169-80 (1993). 3. Wong, D.H., Brodsky, F.M.: 100-kD proteins of Golgi- and trans-Golgi network-associated coated vesicles have related but distinct membrane binding properties. J. Cell. Biol. 117: 1171-9 (1992). 4. Beck, K.A., et al.: Clathrin assembly protein AP-2 induces aggregation of membrane vesicles: a possible role for AP-2 in endosome formation. J. Cell. Biol. 119: 787-96 (1992).

Additional Info

Detects assembly polypeptide 2 (AP2). Recognizes the products of both alpha-adaptin genes, alpha A and alpha C as well as an alternatively spliced isoform of alpha A found in neurons. Species Crossreactivity: Human, Rat, Mouse, Bovine and Primate.