モノクロナール抗γ-アダプチン マウス宿主抗体

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.

ウシ脳由来AP-1アダプタ-。

イムノブロッティングにおいて、ヒトゴルジアダプタ-複合体AP-1の104kDaポリペプチドと反応する抗体です。ウシ肝臓、ヒト心臓繊維芽細胞や培養Madin-Darby bovine kidney(MDBK)細胞に存在する約100kDaのポリペプチドと反応しますが、これらの由来を含め神経芽細胞腫や星状細胞にある110~115kDaの成分とは反応しません。このことから、110kDaおよび115kDaのポリペプチドが、ある種の脳細胞にのみ生じる特異的なバリアントである可能性が考えられます。この抗体は、齧歯類(ラットおよびマウス)のγ-サブユニットは認識しません。MDBK細胞、ヒト心臓繊維芽細胞やアフリカミドリザル腎臓細胞(Vero)を用いた免疫蛍光染色において、抗体の結合は、その大部分がトランスゴルジ網に限定されるとみられています。この抗体を利用してブレフェルジンAの効果が研究されており、ブレフェルジンAは、トランスゴルジ網から出芽するクラスリン被覆小胞に関連するコ-トタンパク質の急速な再配分を引き起こす物質であるといわれています。抗体はゴルジアダプタ-複合体AP-1の免疫アフィニティ-精製にも使用することができます。

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)

Immunofluorescence of Hela cells was performed using monoclonal anti-AP1 (clone 100/3) as the primary antibody.

Mouse monoclonal anti-gamma-adaptin antibody can be used for western blot (1:100, using bovine brain extract), immunoprecipitation and electron microscopy applications.

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).

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