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8606
Actin Nucleation and Polymerization Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Actin Nucleation and Polymerization Antibody Sampler Kit #8606

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Actin Nucleation and Polymerization Antibody Sampler Kit: Image 1

Western blot analysis of A431 cell extracts, untreated (-) or EGF treated (+), using Phospho-Rac1/cdc42 (Ser71) Antibody (upper) or Rac1/Cdc42 Antibody #4651 (lower).

Actin Nucleation and Polymerization Antibody Sampler Kit: Image 2

Western blot analysis of extracts from various cell lines using Rac1/Cdc42 Antibody.

Actin Nucleation and Polymerization Antibody Sampler Kit: Image 3

Confocal immunofluorescent analysis of HeLa cells, serum-starved (left) or EGF-treated #9908 (right), using Wave-2 (D2C8) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Actin Nucleation and Polymerization Antibody Sampler Kit: Image 4

Western blot analysis of extracts from various cell types using Profilin-1 (C56B8) Rabbit mAb.

Actin Nucleation and Polymerization Antibody Sampler Kit: Image 5

Western blot analysis of extracts from various cell types using ARP2 Antibody.

Actin Nucleation and Polymerization Antibody Sampler Kit: Image 6

Western blot analysis of extracts from HeLa, Jurkat and COS cells using ARP3 Antibody.

Actin Nucleation and Polymerization Antibody Sampler Kit: Image 7

Immunoprecipitation of N-WASP from MCF-7 cells, using N-WASP (30D10) Rabbit mAb. Western blot was performed using the same antibody.

Actin Nucleation and Polymerization Antibody Sampler Kit: Image 8

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.

Actin Nucleation and Polymerization Antibody Sampler Kit: Image 9

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 and Polymerization Antibody Sampler Kit: Image 10

Western blot analysis of extracts from various cell types, using N-WASP (30D10) Rabbit mAb.

Actin Nucleation and Polymerization Antibody Sampler Kit: Image 11

Western blot analysis of extracts from various cell types using WAVE-2 (D2C8) XP® Rabbit mAb.

To Purchase # 8606T
Product # Size Price
8606T
1 Kit  (7 x 20 µl) $ 500

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-Rac1/cdc42 (Ser71) Antibody 2461 20 µl
  • WB
H 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
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 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Actin Nucleation and Polymerization Antibody Sampler Kit provides an economical means to evaluate the presence and status of actin nucleation and polymerization. The kit contains enough primary antibody to perform two western blots per primary.

Specificity / Sensitivity

ARP2 Antibody recognizes endogenous levels of total ARP2 protein. ARP3 Antibody recognizes endogenous levels of total ARP3 protein. This antibody does not cross-react with endogenous ARP2. Profilin-1 (C56B8) Rabbit mAb recognizes endogenous levels of total profilin-1 protein. This antibody may cross-react with profilin-2. Phospho-Rac1/cdc42 (Ser71) Antibody recognizes endogenous levels of Rac1/cdc42 only when phosphorylated at serine 71. This antibody may also detect phospho-RhoA (Ser73). Rac1/Cdc42 Antibody recognizes endogenous levels of total rac1 and cdc42 proteins. Based on sequence similarities, this antibody may also detect rac2 and rac3; this antibody does not cross-react with RhoA, RhoB, or RhoC. N-WASP (30D10) Rabbit mAb recognizes endogenous levels of total N-WASP protein. This antibody does not cross-react wth the hematopietic protein, WASP. WAVE-2 (D2C8) XP® Rabbit mAb recognizes endogenous levels of total WAVE-2 protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human profilin-1 protein, the sequence of human N-WASP protein, or central residues of human WAVE-2 protein.

Activation state specific polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser71 of human Rac1/cdc42 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino termini of human rac1 and cdc42 proteins, the sequence of human ARP2 protein, or the amino terminus of human ARP3 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

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).

  1. Wennerberg, K. and Der, C.J. (2004) J Cell Sci 117, 1301-12.
  2. Bernards, A. and Settleman, J. (2004) Trends Cell Biol 14, 377-85.
  3. Rossman, K.L. et al. (2005) Nat Rev Mol Cell Biol 6, 167-80.
  4. Millard, T.H. et al. (2004) Biochem J 380, 1-17.
  5. Suetsugu, S. et al. (1999) Biochem Biophys Res Commun 260, 296-302.
  6. Goley, E.D. and Welch, M.D. (2006) Nat Rev Mol Cell Biol 7, 713-26.
  7. Witke, W. (2004) Trends Cell Biol 14, 461-9.
  8. Boukhelifa, M. et al. (2006) FEBS J 273, 26-33.
  9. Gareus, R. et al. (2006) J Biol Chem 281, 2803-11.
  10. Reinhard, M. et al. (1995) EMBO J 14, 1583-9.
  11. Suetsugu, S. et al. (1998) EMBO J 17, 6516-26.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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