Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Actin Nucleation Antibody Sampler Kit #8355

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
ARP2 Antibody #3128 40 µl W H M R Hm Mk Dm 44 Rabbit
ARP3 Antibody #4738 40 µl W H Mk 47 Rabbit
N-WASP (30D10) Rabbit mAb #4848 40 µl W IP H M R Hm Mk B 65 Rabbit IgG
Diap1 Antibody #5486 40 µl W H Mk 150 Rabbit
Diap2 Antibody #5474 40 µl W IP H M R Mk 130 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Dm=D. melanogaster  B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

ARP2 Antibody detects endogenous levels of total ARP2 protein. ARP3 Antibody detects endogenous levels of total ARP3 protein and does not cross-react with endogenous levels of total ARP2 protein. Diap1 and Diap2 Antibodies recognize endogenous levels of total Diap1 and Diap2 protein, respectively. N-WASP (30D10) Rabbit mAb recognizes endogenous levels of total N-WASP protein and does not cross-react with the hematopoietic protein, WASP.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using ARP2 Antibody #3128.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using ARP3 Antibody #4738.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using N-WASP (30D10) Rabbit mAb #4848.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Diap2 Antibody #5474.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Diap1 Antibody #5486.

Description

The Actin Nucleation Antibody Sampler Kit provides an economical means to evaluate the presence and status of actin nucleation. The kit contains enough primary and secondary antibodies to perform four western blot experiments per primary antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human ARP2, the amino terminus of human ARP3, residues near the carboxy terminus of human diap1 protein, or residues near the amino terminus of human diap2 protein. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding the sequence of human N-WASP protein.

Background

Actin nucleation, the formation of new actin filaments from existing filaments, affects actin filament structure during cell motility, division, and intracellular trafficking. An important actin nucleation protein complex is the highly conserved ARP2/3 complex, consisting of ARP2, ARP3, and ARPC1-5. The ARP2/3 complex promotes branching of an existing actin filament and formation of a daughter filament following activation by nucleation-promoting factors, such as WASP/WAVE or cortactin (1). The formation of podosomes, small cellular projections that degrade the extracellular matrix, is enhanced by ARP2/3 complex action. ARP2/3 competes with caldesmon, an actin binding protein shown to negatively affect podosome formation (2). Along with N-WASP, the ARP2/3 complex regulates nuclear actin filament nucleation and controls actin polymerization during transcription (3).

Formins are a family of large multidomain actin nucleation/polymerization proteins characterized by their catalytic FH2 domains. The mammalian diaphanous-related formin (mDia/diap) subfamily, including mDia1/diap1, mDia2/diap3 and mDia3/diap2, are effectors of Rho family small GTPases. In response to Rho, mDia/diap proteins are involved in the regulation of multiple cell functions including cytoskeletal dynamics, migration, adhesion, polarity, and cell shape (reviewed in 4,5).mDia1/diap1 is activated by GTP-bound Rho, leading to Rho-associated kinase (ROCK)-dependent stress fiber formation (6,7). Rho activation of mDia1 has also been shown to regulate serum response factor (SRF)-dependent transcription (8), and has been implicated in human cancer phenotypes such as ras-mediated transformation, metastasis, and invasion (reviewed in 9).mDia3/diap2, activated by the Rho family small GTPase cdc42, regulates the attachment of microtubules to the kinetochore during mitosis in mammalian cells (10).Rho-dependent activation of mDia2/diap3 is important in assembly of the contractile ring during cytokinesis (11,12).

  1. Goley, E.D. and Welch, M.D. (2006) Nat. Rev. Mol. Cell Biol. 7, 713-726.
  2. Morita, T. et al. (2007) J. Biol. Chem. 282, 8454-8463.
  3. Yoo, Y. et al. (2007) J. Biol. Chem. 282, 7616-7623.
  4. Schönichen, A. and Geyer, M. (2010) Biochim Biophys Acta 1803, 152-63.
  5. Chesarone, M.A. et al. (2010) Nat Rev Mol Cell Biol 11, 62-74.
  6. Watanabe, N. et al. (1999) Nat Cell Biol 1, 136-43.
  7. Ishizaki, T. et al. (2001) Nat Cell Biol 3, 8-14.
  8. Copeland, J.W. and Treisman, R. (2002) Mol Biol Cell 13, 4088-99.
  9. Narumiya, S. et al. (2009) Cancer Metastasis Rev 28, 65-76.
  10. Yasuda, S. et al. (2004) Nature 428, 767-71.
  11. Watanabe, S. et al. (2010) Mol Biol Cell 21, 3193-204.
  12. Watanabe, S. et al. (2008) Mol Biol Cell 19, 2328-38.

Application References

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Protocols

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

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