#8606 Actin Nucleation and Polymerization Antibody Sampler Kit
|Phospho-Rac1/cdc42 (Ser71) Antibody #2461||20 µl||WB||H||M, R||28||Rabbit|
|Rac1/Cdc42 Antibody #4651||20 µl||WB||H, M, R, Mk||21||Rabbit|
|WAVE-2 (D2C8) XP® Rabbit mAb #3659||20 µl||WB, IP, IF-IC||H, M, R, Mk||80||Rabbit IgG|
|Profilin-1 (C56B8) Rabbit mAb #3246||20 µl||WB||H, M, R, Hm, Mk, Dg||15||Rabbit IgG|
|ARP2 Antibody #3128||20 µl||WB||H, M, R, Hm, Mk, Dm||44||Rabbit|
|ARP3 Antibody #4738||20 µl||WB||H, Mk||47||Rabbit|
|N-WASP (30D10) Rabbit mAb #4848||20 µl||WB, IP||H, M, R, Hm, Mk, B||65||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||WB||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey, Hm=Hamster, Dg=Dog, Dm=D. melanogaster, B=Bovine
Western blot analysis of A431 cell extracts, untreated (-) or EGF treated (+), using Phospho-Rac1/cdc42 (Ser71) Antibody (upper) or Rac1/Cdc42 Antibody #4651 (lower).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from various cell types, using N-WASP (30D10) Rabbit mAb.
Western blot analysis of extracts from HeLa, Jurkat and COS cells using ARP3 Antibody.
Western blot analysis of extracts from various cell types using Profilin-1 (C56B8) Rabbit mAb.
Western blot analysis of extracts from various cell types using WAVE-2 (D2C8) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using Rac1/Cdc42 Antibody.
Immunoprecipitation of N-WASP from MCF-7 cells, using N-WASP (30D10) Rabbit mAb. Western blot was performed using the same antibody.
Immunoprecipitation of WAVE-2 from HeLa cells using WAVE-2 (D2C8) XP® Rabbit mAb. Western blot was performed using the same antibody. Lane 1 is 5% input.
Actin nucleation is the process of forming new actin filaments and is necessary to stimulate actin polymerization. Actin polymerization is vital for cell motility, cell division, and cell adhesion. Rac and Cdc42, members of the Rho-GTPase family, play key roles in actin dynamics, membrane trafficking, transcriptional regulation, cell growth, and development (1). GTP binding stimulates the activity of Rac/Cdc42, and the hydrolysis of GTP to GDP through the intrinsic GTPase activity or Rac/Cdc42, rendering it inactive. GTP hydrolysis is aided by GTPase activating proteins (GAPs), while exchange of GDP for GTP is facilitated by guanine nucleotide exchange factors (GEFs). Another level of regulation is achieved through binding of RhoGDI, a guanine dissociation inhibitor, which retains Rho family GTPases, including Rac and Cdc42, in their inactive GDP-bound state (2,3). Hematopoietic WASP and ubiquitously expressed N-WASP are autoinhibited in unstimulated cells. Upon stimulation they are activated by Cdc42, which relieves the autoinhibition in conjunction with phosphatidyl inositol 4,5-bisphosphate (4). Three WAVE (Wasf, SCAR) family proteins are similar in sequence to WASP and N-WASP, but lack the WASP/N-WASP autoinhibition domains and are indirectly activated by Rac (4). WAVE-2 is widely distributed, while WAVE-1 and WAVE-3 are strongly expressed in the brain (5). The highly conserved ARP2/3 complex is an important actin nucleation protein complex consisting of ARP2, ARP3, and ARPC1-5. The ARP2/3 complex promotes branching of existing actin filaments and formation of daughter filaments following activation by nucleation-promoting factors, such as WASP/WAVE or cortactin (6). Profilins are conserved actin binding proteins that affect the rate of actin polymerization by binding actin monomers and promoting exchange of ADP for ATP (reviewed in 7). Profilins bind to proteins involved in the regulation of actin dynamics including paladin (8), dynamin-I (9), VASP (10), and N-WASP (11).
- Wennerberg, K. and Der, C.J. (2004) J Cell Sci 117, 1301-12.
- Bernards, A. and Settleman, J. (2004) Trends Cell Biol 14, 377-85.
- Rossman, K.L. et al. (2005) Nat Rev Mol Cell Biol 6, 167-80.
- Millard, T.H. et al. (2004) Biochem J 380, 1-17.
- Suetsugu, S. et al. (1999) Biochem Biophys Res Commun 260, 296-302.
- Goley, E.D. and Welch, M.D. (2006) Nat Rev Mol Cell Biol 7, 713-26.
- Witke, W. (2004) Trends Cell Biol 14, 461-9.
- Boukhelifa, M. et al. (2006) FEBS J 273, 26-33.
- Gareus, R. et al. (2006) J Biol Chem 281, 2803-11.
- Reinhard, M. et al. (1995) EMBO J 14, 1583-9.
- Suetsugu, S. et al. (1998) EMBO J 17, 6516-26.
DRAQ5 is a registered trademark of Biostatus Limited.
XP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.