ELISA, Immunohistochemistry, Immunoprecipitation, Western Blot. Anti-Osteopontin has been tested for western blotting, immunohistochemistry (formalin-fixed paraffin-embedded sections) and ELISA. The antibody exclusively recognizes C-terminal fragments of both thrombin and MMP-cleaved OPN. The antibody recognizes the full-length osteopontin protein (which runs at 66 kDa on westerns), and 32 kDa for the MMP-cleaved C-fragment, but the 40 kDa N-terminal fragment is not recognized. A 1:1000 dilution will detect strongly approximately 250 ng of OPN protein on a blot. A 1:100-1:300 dilution used for IHC on human breast tumor tissues. No pretreatment is required for IHC when formalin-fixed paraffin-embedded tissue is stained. Specific conditions for reactivity and signal detection should be optimized by the end user.
Reviews / Ratings
If you have used this antibody, please help fellow researchers by submitting reviews to pAbmAbs and antYbuddY.
Description
Osteopontin is directed against human osteopontin. The antibody recognizes the full-length osteopontin protein (which runs at 66 kDa on westerns), as well as the C-terminal fragments of both thrombin and MMP-cleaved OPN. The 32 kDa MMP-cleaved C-fragment is recognized, but not the 40 kDa N-terminal fragment. Reactivity is reported to occur with osteopontin from swine, dog, mouse and rat.
Immunogen
This whole rabbit serum was prepared by repeated immunizations with a synthetic peptide, from the human osteopontin protein, conjugated to KLH using maleimide.
Kim SJ, Shin TK. Immunohistochemical study of osteopontin in boar testis. J Vet Sci. 2007 June; 8(2): 107-110. Published online 2007 June 30. doi: 10.4142/jvs.2007.8.2.107 PMCID: PMC2872706Robertson BW, Bonsal L, Chellaiah MA. Regulation of Erk1/2 activation by osteopontin in PC3 human prostate cancer cells. Mol. Cancer. 2010;9:260. Robertson, B. W., & Chellaiah, M. A. (2010). Osteopontin induces ?-catenin signaling through activation of Akt in prostate cancer cells. Experimental cell research, 316(1), 1-11.Cherepanova O, Gomez D, Shankman L, Swiatlowska P, Williams J, Sarmento O, Alencar G, Hess D, Bevard M, Greene E, Murgai M, Turner S, Geng Y, Bekiranov S, Connelly J, Tomilin A, Owens G. (2016) Activation of the pluripotency factor OCT4 in smooth muscle cells is atheroprotective. Nature Medicine volume 22, pages 657-665.