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Technical Data Sheet

AK6390

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Arp2/3 Complex Regulation
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
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
WP2601 Uenishi, E. et al. (2013) J of Bio Chem 288(36): 25851. WB: MIN6-K8 ?-cells
AP3861 Spillane, M. et al. (2012) J Neurosci. 32(49):17671. ICC: Chicken embryo neuron
CP2621 Williams, H.C. et al. (2012) Circ Res. 111(1):56. ICC, WB: mouse carotid artery,smooth muscle
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
WP2101 Cheney, L. et al. (2011) J Infectious Diseases. 203:1824. WB: mouse adipocytes
WP2601 Cheney, L. et al. (2011) J Infectious Diseases. 203:1824. WB: mouse adipocytes
AP3861 Kalwa, H. & Michel, T. (2011) J Biol Chem. 286(3):2320. WB: bovine aortic endothelial cells
WP2601 Pichot, C.S. et al. (2009) British J Cancer. 1 –10. WB: breast cancer cell lines
Kit Summary
The Arp2/3 complex regulation antibody sampler kit can be used to examine phosphorylation of the Arp2/3 complex subunit Arp2, as well as the Arp2/3 complex regulators N-WASP/WASP and Coronin-1B. The kit contains antibodies to monitor total expression levels of each of these proteins relative to site-specific phosphorylation.
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Western blot of human A431 cells treated with Calyculin A (100 nM) for 30 min. Blot lanes were untreated (lanes 1 & 3) or treated with lambda phosphatase (lanes 2 & 4) then probed with anti-Arp2 (C-terminal) (lanes 1 & 2) or anti-Arp2 (T237/T238) (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. These phosphorylation events may regulate binding to the pointed end of actin filaments, and alanine substitutions of these Arp2 phosphosites inhibit membrane protrusions. Thus, phosphorylation may be another mode of Arp2/3 complex regulation in addition to the activity of nucleation-promoting factors.

Background References
Kelleher, J.F. et al. (1995). J Cell Biol. 131(2):385.
Baba, Y. et al. (1999) Blood 93:2003.
Higgs, H.N. & Pollard, T.D. (2001) Annu Rev Biochem 70:649-676.
Torres, E. & Rosen, M.K. (2003) Mol Cell 11:1215.
Cory, G.O. et al. (2003) Mol Cell. 11(5):1229-39.
Wu, X. et al. (2004) J Biol Chem 279(10):9565.
Foger, N. et al. (2006) Science 313:839.
Uetrecht, A.C. & Bear, J.E. (2006) Trends Cell Biol. 16(8):421.
Cai, L. et al. (2007) Cell 128:915.
LeClaire, L.L. et al (2008). J Cell Biol. 182(4):647.
Soderling, S.H. (2009). Sci Signal. 2(55):pe5 (Review).
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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