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. Author manuscript; available in PMC: 2011 Mar 1.
Published in final edited form as: Int J Oral Maxillofac Surg. 2010 Jan 25;39(3):272–281. doi: 10.1016/j.ijom.2009.12.017

Fig. 3.

Fig. 3

Western immunoblotting for DSPP and its fragments, DMP1 and its fragments, BSP and OPN. (A) Western immunoblotting using anti-DSP polyclonal antibody for fractions 27–39 of Q-Sepharaose chromatography of NCPs extracted from rat condylar cartilage. Positive control (Cont): 0.3 μg of DSP isolated from rat incisor dentin; 27–39: 60 μl of sample from fractions 27-39 were treated with 2.5% of β-mercaptoethanol before loading. Long arrow indicates the migrating position of full-length DSPP; arrowhead indicates the migrating position of DSP. (B) Western immunoblotting using anti-DMP1-N-9B6.3 monoclonal antibody. Cont: 1 μg of the NH2-terminal (37 kDa) fragment of DMP1 and the full-length form of DMP1 isolated from rat long bone. 51: 60 μl of sample from fraction 51 of Q-Sepharaose chromatography of NCPs extracted from rat condylar cartilage. Both the 37 kDa fragment (arrowhead) and full-length form of DMP1 (long arrow) are detected in the extract from the rat condylar cartilage. In comparison with the DMP1 molecular species in the extract from rat long bone (Cont in Fig. 3B), the full-length form of DMP1 is more abundant in the extract from the cartilage. (C) Western immunoblotting using anti-DMP1-C-857 polyclonal antibody. Cont: 1 μg of the COOH-terminal (57 kDa) fragment and the full-length form of DMP1 isolated from rat long bone. 51: 60 μl of sample from fraction 51 of Q-Sepharaose chromatography of NCPs extracted from rat condylar cartilage. While the full-length form of DMP1 (long arrow) was visualized, the 57 kDa fragment (arrowhead) was hardly detectable in the extract from the rat condylar cartilage. (D) Western immunoblotting using anti-BSP-10D9.3 monoclonal antibody. Cont: 1 μg of BSP isolated from rat long bone. 69: fraction 69 of Q-Sepharaose chromatography of NCPs extracted from rat condylar cartilage. Arrow indicates the migrating position of BSP. (E) Western immunoblotting using the anti-OPN polyclonal antibody. Cont: 1 μg of OPN isolated from rat long bone. 51: fraction 51 of Q-Sepharaose chromatography of NCPs extracted from rat condylar cartilage. Arrow indicates the migrating position of OPN.