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Technical Data Sheet


Phospho-Serine, -Threonine, & -Tyrosine
Immunocytochemistry Kit


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
PM3801 Osma-Garcia, I.C. et al. (2015) Eur J Immunol. doi: 10.1002 WB: mouse macrophages
PP2221 Hamada-Kawaguchi, N. et al. (2015) PLoS One. 10(3):e0121484 ICC: fly ovary
PP2551 Kommaddi, R.P. et al. (2015) J Biol Chem. 3;290(14):8888 WB: COS-7 cells
PP2551 Tsai, C.F. et al. (2015) J Agric Food Chem. 9;63(48):10388 WB: Huh7 cells
PM3801 Fester, L. et al. (2015) JSBMB S0960-0760(15)30109. WB: Hippocampal neurons
PP2221 Sroyraya, M. et al. (2013) Microsc Res Tech. 76(1):103. WB: Swimming Crab
PP2551 Kim, S. et al. (2013) Diabetes. 62(2):471. WB, IP: mouse adipocytes
PP2551 Sroyraya, M. et al. (2013) Microsc Res Tech. 76(1):103. WB: Swimming Crab
PM3801 Morinaga, J. et al. (2013) AJP Renal Phys. 305(2)F173. WB: Normal Rat Kidney Fibroblast
PP2551 Hummel, S. et al. (2012) Appl Environ Microbiol. 78(4):1140. WB: human colonic adenocarcinoma cells
PP2551 Laluk, K. et al. (2011) Plant Cell 23(8):2831. WB: Arabidopsis BIK1 and Myelin basis protein
Kit Summary
The phospho-serine, -threonine, and -tyrosine kit can be used to examine the pattern of phosphoserine-threonine relative to phosphotyrosine, or relative to a target protein. The kit includes monoclonal and polyclonal antibodies to phosphoserine-threonine and phosphotyrosine along with Goat-anti-Rabbit conjugated to DyLight® 488 and Goat anti-Mouse conjugated to DyLight® 594 for dual labeling experiments.

Immunocytochemical labeling of phosphorylation in A431 treated with calyculin A (left panel) or pervanadate (right panel). The fixed cells were labeled with anti-Phosphoserine/threonine (PM3801) and anti-Phosphoserine/threonine (PP2551) (left panel) or with anti-Phosphotyrosine (PM3751) and anti-Phosphotyrosine (PP2221) (right panel). The antibodies were detected using appropriate secondary antibody conjugated to DyLight® 488 or DyLight® 594.

Phosphorylation of specific tyrosine, serine, and threonine residues is an important post-translational modification for regulating the activity of proteins. Stimulation of a variety of cell signaling pathways activates the receptor and non-receptor kinases that mediate these protein modifications. Antibodies that can detect phosphotyrosine, phosphoserine, or phosphothreonine residues are excellent tools for characterizing changes in the post-translational state of a broad range of phospho-proteins. Immunoprecipitation of proteins of interest, followed by detection of phosphorylation using anti-phosphotyrosine or anti-phosphoserine/threonine antibody is commonly used to correlate changes in phosphorylation state with alterations in protein activity. Immunocytochemistry can also be used to examine changes in the level and localization of phosphotyrosine, phosphoserine, or phosphothreonine in cells after drug stimulation or during specific cell states.

Background References
Ross, A.H. et al. (1981) Nature 294:654.
Hunter T.(1987) Cell. 50(6):823.
Wang, J.Y.J. (1988) Anal. Biochem 172:1.
Krishna, R.G. & Wold, F. (1993) Adv Enzymol Rel Areas Mol Biol 67:265.
Yaffe, M.B. & Elia, A.E. (2001) Curr Opin Cell Biol 13, 131-8.
Buffer and Storage
Mouse monoclonal, rabbit polyclonal, and secondary reagents are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
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