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Technical Data Sheet

AK6670

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Arp2/3 Complex
Antibody Sampler Kit
Price

$395

Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
AP4351 Kim, Y.B. et al. (2015) J Biol Chem. 290(34):21076 WB: PC3 cells
AP3861 Osma-Garcia, I.C. et al. (2015) Eur J Immunol. doi: 10.1002 WB: mouse macrophages
AP3871 Osma-Garcia, I.C. et al. (2015) Eur J Immunol. doi: 10.1002 WB: mouse macrophages
AP4321 Ho, H. et al. (2013) Pigment Cell Melanoma Res 26(2):218. WB: human A375 melanoma
AP4351 Ho, H. et al. (2013) Pigment Cell Melanoma Res 26(2):218. WB: human A375 melanoma
AP3861 Spillane, M. et al. (2012) J Neurosci. 32(49):17671. ICC: Chicken embryo neuron
AP3861 Kalwa, H. & Michel, T. (2011) J Biol Chem. 286(3):2320. WB: bovine aortic endothelial cells
AP3861 Spillane, M. et al. (2011) Dev Neurobiol. 71(9):747. ICC: chick dorsal root ganglia
AP3871 Kalwa, H. & Michel, T. (2011) J Biol Chem. 286(3):2320. WB: bovine aortic endothelial cells
Kit Summary
The Arp2/3 complex kit can be used to detect expression levels of Arp2, Arp3, and Arpc1b in many different species. In addition, phospho-specific antibodies are included to detect phosphorylation of Arp2 (Thr-237/Thr-238) and Arpc1b (Thr-21).
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Western blot of Arpc1b phosphorylation in A431 stimulated with calyculin A (100 nM) for 30 min (lanes 1 & 3). The blots were treated with lambda phosphatase to remove phosphorylation (lanes 2 & 4), then probed with anti-Arpc1b (C-terminal region) (lanes 1 & 2) or anti-Arpc1b (Thr-21) (lanes 3 & 4).

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Immunocytochemical labeling of Arp2 phosphorylation in rat PC12 cells differentiated with NGF. The cells were probed with Arp2 (C-terminal region) and Arp2 (Thr-237/Thr-238) rabbit polyclonal antibodies, then the antibodies were detected using appropriate secondary antibody conjugated to Cy3.

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Background
Cellular morphology, adhesion, and motility occur through dynamic reorganization of actin-based superstructures. Actin-binding proteins are critical for regulating actin polymerization and superstructure formation. The Arp2/3 complex is an actin polymerization-inducing complex that includes Arp2, Arp3, p41-Arc, p34-Arc, p21-Arc, p20-Arc, and p16-Arc. Several nucleation promoting factors, such as WASP and coronin, regulate the activity of the Arp2/3 complex. In addition, the Arp2/3 complex may be regulated by phosphorylation of specific subunits in the complex. Arp2 has two phosphosites, Thr-237 and Thr-238, that are evolutionarily conserved, and are phosphorylated along with Tyr-202 in response to growth factor stimulation. p41-Arc (Arpc1b) subunit Arpc1 has two isoforms in humans, Arpc1a and Arpc1b. PAK1 can bind and phosphorylate Thr-21 in Arpc1b leading to growth factor-stimulated cell motility. In addition, Arpc1b colocalizes with γ-tubulin at centrosomes and stimulates Aurora A activity.

Background References
Kelleher, J.F. et al. (1995). J Cell Biol. 131(2):385.
Welch, M.D. et al. (1997) J Cell Biol. 138(2):375.
Vadlamudi, R.K. et al. (2004) EMBO Rep. 5(2):154.
LeClaire, L.L. et al (2008). J Cell Biol. 182(4):647.
Soderling, S.H. (2009). Sci Signal. 2(55):pe5 (Review).
Molli, P.R. et al. (2010) J Cell Biol. 190(1):101.
Buffer and Storage
Rabbit polyclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
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