Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

EEA1 (C45B10) Rabbit mAb #3288

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H M R Endogenous 170 Rabbit IgG

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

Protocols

Specificity / Sensitivity

EEA1 (C45B10) Rabbit mAb detects endogenous levels of total EEA1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser70 of human EEA1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using EEA1 (C45B10) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using EEA1 (C45B10) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

Background

EEA1 is an early endosomal marker and a Rab5 effector protein essential for early endosomal membrane fusion and trafficking (1-2). The carboxy terminus of EEA1 contains a FYVE domain which binds to phosphatidylinositol-3-phosphate (PtdIns(3)P), targeting EEA1 to early endosomes (3). The stable association of EEA1 with the endosomal membrane is regulated by PI3 kinase, Rab5 and calcium/calmodulin (4-6). Once on the membrane, EEA1 interacts with Rab5, NSF and syntaxin 13 to promote early endosomal membrane docking and fusion (7).

  1. Mu, F.T. et al. (1995) J. Biol. Chem. 270, 13503-13511.
  2. Christoforidis, S. et al. (1999) Nature 397, 621-625.
  3. Gaullier, J.M. et al. (1998) Nature 394, 432-433.
  4. Patki, V. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 7326-7330.
  5. Lawe, D.C. et al. (2003) Mol. Biol. Cell 14, 2935-2945.
  6. Simonsen, A. et al. (1998) Nature 394, 494-498.
  7. McBride, H.M. et al. (1999) Cell 98, 377-386.

Application References

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Companion Products

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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