Cell Signaling Technology

Product Pathways - Vesicle Trafficking

Phospho-AP2M1 (Thr156) (D4F3) Rabbit mAb #7399

No. Size Price
7399S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
7399 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 50 Rabbit IgG
IP 1:100
IF-IC 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry),

Homology

Species predicted to react based on 100% sequence homology: Mouse, Rat, Monkey,

Specificity / Sensitivity

Phospho-AP2M1 (Thr156) (D4F3) Rabbit mAb recognizes endogenous levels of AP2M1 protein only when phosphorylated at Thr156.

Phospho-AP2M1 (Thr156) (D4F3)Rabbit mAb可以识别156位苏氨酸被磷酸化的AP2M1蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr156 of human AP2M1 protein.

单克隆抗体由合成肽段免疫动物产生,该肽段与人AP2M1蛋白156位苏氨酸附近氨基酸一致。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or H2O2-treated (4 mM, 15 min; right), using Phospho-AP2M1 (Thr156) (D4F3) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).使用Phospho-AP2M1 (Thr156) (D4F3)Rabbit mAb(绿色)对未处理(左)或用H2O2处理(4mM,30分钟)的HeLa细胞提取物进行激光共聚焦免疫荧光分析。肌动蛋白丝使用DY-554鬼笔环肽标记(红色)。蓝色假色= DRAQ5® #4084 (fluorescent DNA dye).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, serum-starved overnight and either untreated or H2O2-treated (4 mM, 30 min), using Phospho-AP2M1 (Thr156) (D4F3) Rabbit mAb (upper) or β-Actin Antibody #4967 (lower).使用Phospho-AP2M1 (Thr156) (D4F3)Rabbit mAb(上)或β-Actin抗体#4967(下)对血清饥饿过夜及不处理或用H2O2处理(4mM,30分钟)的HeLa细胞提取物进行western blot分析。

Background

The AP-2 coat assembly protein complex is an important component of clathrin-coated pits involved in receptor-mediated endocytosis at the plasma membrane (1-3). Each AP-2 heterotetramer is composed of α, β, μ, and σ protein subunits. The 50 kDa μ subunit (AP-2μ, AP2M1) is located at the core of the AP-2 complex and mediates interaction between the cargo protein and the clathrin-coated pit (1-4). The carboxy-terminal AP2M1 region recognizes the tyrosine-based, endocytotic sorting motif YXXφ found in cargo proteins and helps to bring the cargo protein to the clathrin-coated pit. Non-canonical, tyrosine-based endocytotic sorting signals can also promote interaction between cargo proteins and AP2M1 (5,6). AP2M1 plays an essential role in molecular signaling as it couples receptor-mediated endocytosis and pathways involving membrane receptors (7-9), matrix metalloproteinases (10), and ion channel proteins (11). Phosphorylation of specific AP2M1 residues and binding of lipids to this adaptor protein can regulate AP2M1 activity (12,13). Phosphorylation of AP2M1 at Thr156 by adaptor-associated kinase 1 (AAK1) stimulates affinity binding of AP2M1 to cargo protein signals (14). AP-2盖组装蛋白复合物是网格蛋白盖凸面的重要组成部分,涉及到细胞膜上的受体介导的内吞作用(1-3)。每一个AP-2异四聚体都由α, β, μ, 和σ蛋白亚基组成。50 kDa 的μ亚基(AP-2μ, AP2M1)位于AP-2复合物的核心,介导了复合物和cargo蛋白以及网格蛋白盖凸面的相互作用(1-4)。AP2M1的羧基端可以识别cargo蛋白中的酪氨酸,内吞分类基序YXXφ,协助cargo蛋白到达网格蛋白盖凸面。非经典的,基于酪氨酸的内吞识别信号也可以促进cargo蛋白和AP2M1间的相互作用(5,6)。AP2M1协助受体介导的内吞作用和信号通路,这些通路涉及细胞膜受体(7-9),金属蛋白酶(10),以及离子通道蛋白(11),因此在分子信号中发挥了重要作用。AP2M1特异位点的磷酸化以及与脂质和受体蛋白结合能够调控AP2M1的活性(12,13)。156位苏氨酸被接头相关激酶1 (AAK1)磷酸化会激活AP2M1和cargo蛋白信号的亲和性(14)。

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  2. Ohno, H. et al. (1995) Science 269, 1872-5.
  3. Traub, L.M. (2003) J Cell Biol 163, 203-8.
  4. Boll, W. et al. (1996) EMBO J 15, 5789-95.
  5. Royle, S.J. et al. (2002) J Biol Chem 277, 35378-85.
  6. Royle, S.J. et al. (2005) J Cell Sci 118, 3073-80.
  7. Chin, Y.R. and Horwitz, M.S. (2005) J Virol 79, 13606-17.
  8. Wernick, N.L. et al. (2005) J Biol Chem 280, 7309-16.
  9. Johannessen, L.E. et al. (2006) Mol Cell Biol 26, 389-401.
  10. Uekita, T. et al. (2001) J Cell Biol 155, 1345-56.
  11. Chen, Z. et al. (2006) Am J Respir Cell Mol Biol 35, 127-32.
  12. Höning, S. et al. (2005) Mol Cell 18, 519-31.
  13. Olusanya, O. et al. (2001) Curr Biol 11, 896-900.
  14. Conner, S.D. and Schmid, S.L. (2002) J Cell Biol 156, 921-9.

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