Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Cytoskeletal Signaling

Clathrin Heavy Chain (D3C6) XP® Rabbit mAb #4796

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H M R Mk (Pg) Endogenous 190 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Clathrin Heavy Chain (D3C6) XP® Rabbit mAb detects endogenous levels of total clathrin protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro1663 of human clathrin heavy chain protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Clathrin Heavy Chain (D3C6) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of SH-SY5Y cells using Clathrin Heavy Chain (D3C6) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

Clathrin-coated vesicles provide for the intracellular transport of cargo proteins following endocytosis and during multiple vesicle trafficking pathways. Vesicles form at specialized areas of the cell membrane where clathrin and associated proteins form clathrin-coated pits. Invagination of these cell membrane-associated pits internalizes proteins and forms an intracellular clathrin-coated vesicle (1,2). Clathrin is the most abundant protein in these vesicles and is present as a basic assembly unit called a triskelion. Each clathrin triskelion is composed of three clathrin heavy chains and three clathrin light chains. Clathrin heavy chain proteins are composed of several functional domains, including a carboxy-terminal region that permits interaction with other heavy chain proteins within a triskelion, and a globular amino-terminal region that associates with other vesicle proteins (2). Adaptor proteins, such as AP2, epsin and EPS15, are responsible for the recruitment of vesicle proteins to sites of pit formation and the assembly of the clathrin-coated vesicle. Following vesicle invagination, the GTPase dynamin constricts the neck of the nascent vesicle to complete formation of the free, cytosolic vesicle (3,4).

  1. Rodriguez-Boulan, E. et al. (2005) Nat. Rev. Mol. Cell Biol. 6, 233-247.
  2. Mousavi, S.A. et al. (2004) Biochem. J. 377, 1-16.
  3. Rappoport, J.Z. et al. (2004) Traffic 5, 327-237.
  4. Brett, T.J. and Traub, L.M. (2006) Curr. Opin. Cell Biol. 18, 395-406.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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