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Technical Data Sheet

GK6980

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Girdin 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
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
Kit Summary
The Girdin phospho-regulation antibody sampler kit can be used to detect phosphorylation of Ser-1416, Ser-1674 and Tyr-1764 relative to total Girdin expression levels. The kit includes rabbit polyclonal, phospho-specific antibodies to Ser-1416, Ser-1674, and Tyr-1764. The kit also includes an anti-Rabbit Light Chain specific:HRP secondary reagent for detection of antibodies in Western blot, ELISA, or immunocytochemistry.
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Enlarge

Western blot image of human A431 cell lysates treated with pervanadate (lanes 1-4). The blot was treated with alkaline phosphatase to dephosphorylate Girdin phosphosites (lanes 2 & 4). The blot was probed with mouse monoclonal anti-Girdin (lanes 1 & 2) or rabbit polyclonal anti-Girdin (Tyr-1764), phospho-specific (lanes 3 & 4).

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Enlarge

Western blot image of human A431 cell lysates treated with Calyculin A (lanes 1-4). The blot was treated with lambda phosphatase to dephosphorylate Girdin phosphosites (lanes 2 & 4). The blot was probed with mouse monoclonal anti-Girdin (lanes 1 & 2) or rabbit polyclonal anti-Girdin (Ser-1674), phospho-specific (lanes 3 & 4).

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Background
Girdin, a member of the CCDC88 (Hook related protein) family, is an actin binding protein involved with cell migration and maintaining cytoskeletal organization. Girdin has conserved domains at the N- and C-terminus that bind microtubules and actin, respectively. It enhances PI3-kinase dependent phosphorylation of proteins, most notably Akt. This same activity can contribute to tumor proliferation, invasion, and metastasis in breast, ovarian, prostate, and pancreatic tissues. Girdin is phosphorylated at three separate locations: Ser-1416, Ser-1674, and Tyr-1764. Ser-1416 is the primary Akt phosphorylation site, while Cyclin-dependent kinases interact with Girdin and phosphorylate Ser-1674. Multiple receptor tyrosine kinases can bind girdin and phosphorylate Tyr-1764.

Background References
Simpson, F. (2005) Traffic. 6(6):442
Jiang, P. (2008) Cancer Res. 1;68(5):1310
Bhandari, D. et al. (2009) Mol Biol Cell. 20:1324.
Lin, C. (2011) Sci Signal. 4(192): ra64
Lin, C. et al. (2014) Mol Biol Cell. 25(22):3654
Bhandari, D. et al. (2015)Proc Natl Acad Sci. 112(35):E4874
Hayano S. et al, (2015) J Mol Cell Cardiol. 88:55
Buffer and Storage
Primary antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagent is supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.
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