Revision 1

#8606Store at -20C

1 个试剂盒

(7 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-Rac1/cdc42 (Ser71) Antibody 2461 20 µl 28 kDa Rabbit 
Rac1/Cdc42 Antibody 4651 20 µl 21 kDa Rabbit 
WAVE-2 (D2C8) XP® Rabbit mAb 3659 20 µl 80 kDa Rabbit IgG
Profilin-1 (C56B8) Rabbit mAb 3246 20 µl 15 kDa Rabbit IgG
ARP2 Antibody 3128 20 µl 44 kDa Rabbit 
ARP3 Antibody 4738 20 µl 47 kDa Rabbit 
N-WASP (30D10) Rabbit mAb 4848 20 µl 65 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

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.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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.

Background References

    Trademarks and Patents

    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.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    仅供研究使用。不得用于诊断流程。
    Revision 1
    #8606

    Actin Nucleation and Polymerization Antibody Sampler Kit

    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 1 Expand Image
    使用 Phospho-Rac1/cdc42 (Ser71) Antibody（上图）或 Rac1/Cdc42 Antibody #4651（下图）对未处理的 (-) 或经 EGF 处理的 A431 细胞提取物进行蛋白质印迹分析。
    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 2 Expand Image
    使用 ARP2 Antibody 对不同细胞类型的提取物进行蛋白质印迹分析。
    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 3 Expand Image
    使用 Profilin-1 (C56B8) Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹分析。
    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 4 Expand Image
    使用 WAVE-2 (D2C8) XP® Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹分析。
    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 5 Expand Image
    使用 Rac1/Cdc42 Antibody 对不同细胞系提取物进行蛋白质印迹分析。
    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 6 Expand Image
    使用 ARP3 Antibody 对 HeLa、Jurkat 和 COS 细胞提取物进行蛋白质印迹分析。
    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 7 Expand Image
    使用 N-WASP (30D10) Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹分析。
    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 8 Expand Image
    一抗与靶标蛋白结合之后，与偶联 HRP 的二抗形成复合体。添加 LumiGLO®，在酶催化分解期间发光。
    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 9 Expand Image
    使用 WAVE-2 (D2C8) XP® Rabbit mAb 对 HeLa 细胞中提取的 WAVE-2 进行免疫沉淀。使用同一抗体进行蛋白质进行印迹分析。泳道 1 是 5% 输入对照。
    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 10 Expand Image
    使用 N-WASP (30D10) Rabbit mAb 对 MCF-7 细胞中提取的 N-WASP 进行免疫沉淀。使用同一抗体进行蛋白质进行印迹分析。
    Actin Nucleation and Polymerization Antibody Sampler Kit: Image 11 Expand Image
    使用 Wave-2 (D2C8) XP® Rabbit mAb（绿色）对血清饥饿（左）或经 EGF 处理的 #9908（右）HeLa 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084（DNA 荧光染料）。
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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    仅供研究使用。不得用于诊断流程。