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Technical Data Sheet

CK6180

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Coronin-1B Phospho-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
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
CP2621 Williams, H.C. et al. (2012) Circ Res. 111(1):56. ICC, WB: mouse carotid artery,smooth muscle
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
Kit Summary
The coronin-1B phospho-regulation antibody sampler kit can be used to detect the level of phosphorylation of coronin-1B (Ser-2) relative to the total expression level of coronin-1B. The kit also includes peptides for antibody blocking experiments and a secondary reagent for antibody detection.
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Western blot analysis of human A431 cells treated with Calyculin A (100 nM) for 30 min (lanes 1 & 3) before treatment with lambda phosphatase (lanes 2 & 4). The blots were probed with anti-Coronin-1B (C-terminal region) (lanes 1 & 2) or anti-Coronin-1B (Ser-2) (lanes 3 & 4).

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Immunocytochemical labeling of coronin-1B in rabbit spleen fibroblasts treated with Calyculin A. The cells were labeled with rabbit polyclonal Coronin-1B (C-terminus) and Coronin-1B (Ser-2) antibodies, then detected using appropriate secondary antibodies conjugated to Cy3. The antibodies were used in the absence (left) or presence (right) of their respective blocking peptide (CX2585 or CX2625).

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Background
Coronins are highly-conserved F-actin binding proteins that play important roles in lamellipodial protrusion during various types of cell motility. In yeast, coronins regulate cytoskeletal changes through inhibition of Arp2/3 complex. Human coronins have been classified in three subgroups: type I (coronin-1A, -1B, -1C), type II (coronin-2A, -2B), and type III (coronin-7). These coronins have at least one large β-propeller region that mediates protein-protein interactions and type I and II coronins have coiled-coil regions involved in oligiomerization. Coronin-1B is ubiquitously expressed and localizes to the leading edge of cell protrusions in migrating fibroblasts. Both Coronin-1B and Coronin-1A interaction with Arp2/3 complex may be regulated by phosphorylation. PKC phosphorylates the N-terminus at Ser-2, and this phosphorylation reduces interactions with Arp2/3 leading to diminished cell motility.

Background References
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.
Buffer and Storage
Rabbit polyclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide, and the peptides are supplied in PBS and 0.05% sodium azide. Store all at –20°C. Stable for 1 year.
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