Immunofluorescence of Hela cells was performed using monoclonal anti-AP1 (clone 100/3) as the primary antibody.
Applications in which this antibody has been used successfully, and the associated peer-reviewed papers, are given below.Immuno-electron microscopy (1 paper)Immunofluorescence (1 paper)
Mouse monoclonal anti-gamma-adaptin antibody can be used for western blot (1:100, using bovine brain extract), immunoprecipitation and electron microscopy applications.
In immunofluorescence using MDBK cells, human heart fibroblasts, and African green monkey kidney cells (Vero), the binding of antibody appears to be largely confined to the trans-Golgi network. The antibody has been used for studies on the effects of Brefeldin A, a substance causing rapid redistribution of coat proteins associated with the clathrin-coated vesicles that bud from the trans-Golgi network. The antibody is also useful for the immunoaffinity purification of the Golgi adaptor complex AP-1.
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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. Clathrin-coated membranes at or originating from the plasma membrane contain the AP-2 adaptor, while the AP-1 adaptor appears to be largely restricted to clathrin-coated membranes of the trans-Golgi network. Both the Golgi-associated AP-1 adaptor and the plasma-membrane associated AP-2 adaptor are heterotetrameric protein complexes. AP-1 adaptor consists of four subunits termed b1 (formerly b′, 110 kDa), g (100 kDa), m1 (47 kDa), and s1 (20 kDa). b1 and g subunits (b1 and g adaptins) from neuronal sources behave in standard SDS-PAGE like 115 kDa and 104 kDa polypeptides, respectively.
AP-1 adaptor from bovine brain.
The product is provided as ascites fluid with 0.1% sodium azide as a preservative.
Mouse monoclonal clone 100/3 anti-g-Adaptin antibody reacts in immunoblotting with the 104 kDa polypeptide of the Golgi adaptor complex AP-1. 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 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 does not recognize the γ-subunit in rodents (rat and mouse).