trap staining solution mix  (Millipore)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: FAM210A is a novel determinant of bone and muscle structure and strength

doi: 10.1073/pnas.1719089115

Figure Lengend Snippet: Phenotypic characterization of Fam210a+/− mice. (A) Serum calcium (Ca) and phosphorus (P) in Ct (female, n = 9; male, n = 7) and Fam210a+/− (female, n = 8; male; n = 8) mice, and the level of Fam210a protein in serum and muscle extracts of Ct (n = 5) and Fam210a+/− (n = 5) mice using ELISA. Undetectable levels were less than 0.3 ng/mL. (B) BMD and BMC of Ct (female, n = 9; male, n = 9) and Fam210a+/− (female, n = 10; male, n = 9) mouse bones. (C) Parameters of biomechanical bone strength, i.e., the maximal load, stiffness, and work-to-failure, measured by three-point bending tests, in Ct (female, n = 14; male, n = 11) and Fam210a+/− (female, n = 11; male, n = 12) mouse femurs. (D) Representative 3D reconstructions of μCT images of trabecular and cortical bone in Ct (n = 3) and Fam210a+/− (n = 3) mouse femurs. (E) Representative micrographs of double calcein-labeled sections of trabecular and cortical bone of Ct and Fam210a+/− mice; MAR and BFR in trabecular and cortical bone of Ct (n = 4) and Fam210a+/− (n = 4) mice calculated using calcein double labeling. (F) Oc.N/BS and Oc.S/BS in trabecular and cortical bone of Ct (trabecular bone, n = 8; cortical bone, n = 10) and Fam210a+/− (trabecular bone, n = 8; cortical bone, n = 9) mice using TRAP staining. (G) Grip strength in Ct (female, n = 5; male, n = 5) and Fam210a+/− (female, n = 5; male, n = 5) mice; lean mass of all limbs in Ct (female; n = 4, male; n = 4) and Fam210a+/− (female, n = 5; male, n = 4). Solid bars indicate values in Ct, and hatched bars indicate values in Fam210a+/− mice. Data represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 using a two-sided Student’s t test. Analyses were performed at 56 d of age.

Article Snippet: Briefly, the sections were incubated in TRAP Staining Solution Mix, containing sodium acetate anhydrous (Sigma), l -(+) tartaric acid (Sigma), and glacial acetic acid at 37° for 30 min.

Techniques: Enzyme-linked Immunosorbent Assay, Labeling, Staining

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: FAM210A is a novel determinant of bone and muscle structure and strength

doi: 10.1073/pnas.1719089115

Figure Lengend Snippet: Additional phenotypic characterization of TFam210a−/− mice. (A) Representative 3D reconstructions of μCT images of trabecular and cortical bone of Ct (n = 4) and TFam210a−/− (n = 4) mouse femurs. (B) Representative micrographs of double calcein-labeled sections of trabecular and cortical bone of Ct and TFam210a−/− mice; MAR and BFR in trabecular and cortical bone Ct (n = 4) and TFam210a−/− (n = 4) mice calculated using calcein double labeling. (C) Oc.N/BS and Oc.S/BS in trabecular and cortical bone of Ct (Oc.N/BS: trabecular bone, n = 5; cortical bone, n = 5; Oc.S/BS: trabecular bone, n = 5; cortical bone, n = 5) and TFam210a−/− (Oc.N/BS: trabecular bone, n = 4; cortical bone, n = 5; Oc.S/BS: trabecular bone, n = 5; cortical bone; n = 5) mice using TRAP staining. (D) Grip strength in Ct (female; n = 5, male; n = 5) and TFam210a−/−, (female; n = 4, male; n = 4) mice; lean mass of all limbs in Ct (female; n = 5, male; n = 3) and TFam210a−/− (female; n = 4, male; n = 4). Solid bars indicate values in Ct, and hatched bars indicate values in TFam210a−/− mice. Data represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 using a two-sided Student’s t test.

Article Snippet: Briefly, the sections were incubated in TRAP Staining Solution Mix, containing sodium acetate anhydrous (Sigma), l -(+) tartaric acid (Sigma), and glacial acetic acid at 37° for 30 min.

Techniques: Labeling, Staining

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: FAM210A is a novel determinant of bone and muscle structure and strength

doi: 10.1073/pnas.1719089115

Figure Lengend Snippet: Additional phenotypic characterization of TFam210aMus−/− mice. (A) Representative 3D reconstructions of μCT images of trabecular and cortical bone in Ct (n = 4) and TFam210aMus−/− (n = 4) mouse femurs. (B) Representative micrographs of double calcein-labeled sections of trabecular and cortical bone of Ct and TFam210aMus−/− mice; MAR and BFR in trabecular and cortical bone Ct (n = 4) and TFam210aMus−/− (n = 4) mice calculated using double calcein labeling. (C) Oc.N/BS and Oc.S/BS in trabecular and cortical bone of Ct (Oc.N/BS: trabecular bone, n = 5; cortical bone, n = 5; Oc.S/BS: trabecular bone, n = 5; cortical bone, n = 5) and TFam210aMus−/− (Oc.N/BS: trabecular bone, n = 5; cortical bone, n = 5; Oc.S/BS: trabecular bone, n = 5; cortical bone; n = 5) mice using TRAP staining. Solid bars indicate values in Ct, and hatched bars indicate values in TFam210aMus−/− mice. (D) RT-PCR to assess the expression of troponin c2, Tnnc2, Cdh15, Myog, and Mmp12 in primary muscle cells from Ct (n = 4) and TFam210aMus−/− (n = 4) mice. Data are expressed relative to the GAPDH mRNA value. Data represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 using a two-sided Student’s t test.

Article Snippet: Briefly, the sections were incubated in TRAP Staining Solution Mix, containing sodium acetate anhydrous (Sigma), l -(+) tartaric acid (Sigma), and glacial acetic acid at 37° for 30 min.

Techniques: Labeling, Staining, Reverse Transcription Polymerase Chain Reaction, Expressing