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Technical Data Sheet

GP5781

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Girdin (Ser-1674), phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
250 kDa

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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).

Application
Dilution


ELISA
1:1000


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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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
Hayano S. et al, (2015) J Mol Cell Cardiol. 88:55
Immunogen
Phospho-Girdin (Ser-1674) synthetic peptide (coupled to carrier) corresponding to amino acid residues around serine 1674 in human Girdin. This sequence has high homology to rat and mouse Girdin, and has low homology to similar regions in other Girdin family members.
Buffer and Storage
Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
The antibody was cross-adsorbed to unphosphorylated Girdin (Ser-1674) peptide before affinity purification using phospho-Girdin (Ser-1674) peptide. This antibody detects a 250 kDa* protein on SDS-PAGE immunoblots of human A431 cells stimulated with Calyculin A. This reactivity is not observed after lambda phosphatase treatment to dephosphorylate Girdin.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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