rankl murine srank ligand mini abts elisa development kit  (PeproTech)

Journal: Brazilian oral research

Article Title: Different engagement of TLR2 and TLR4 in Porphyromonas gingivalis vs. ligature-induced periodontal bone loss

doi: 10.1590/1807-3107BOR-2017.vol31.0063

Figure Lengend Snippet: Gingival RANKL mRNA and protein expression levels of P. gingivalis-induced and ligation-induced experimental periodontitis in WT, TLR2 KO, TLR4 KO, and TLR2&4 KO mice. Gingival tissues on the palatal side were collected under a surgical microscope from maxillae and then homogenized for RNA extraction or protein measurement. Gingival RANKL mRNA levels were determined by real-time PCR in the control group, the P. gingivalis infection group, and the ligation group of WT mice (A), TLR2 KO mice (B), TLR4 KO mice (C), and TLR2&4 KO mice (D) (means ± SE, n = 5 mice per group, *p < 0.05, **p < 0.01). Gingival RANKL protein expression levels were measured by ELISA in the control group, the P. gingivalis infection group, and the ligation group of WT mice (E), TLR2 KO mice (F), TLR4 KO mice (G), and TLR2&4 KO mice (H) (means ± SE, n = 5 mice per group, *p < 0.05, **p < 0.01).

Article Snippet: The secreted RANKL level in the gingival homogenate was detected by means of a murine RANKL ELISA development kit (PeproTech, Rocky Hill, USA) following the manufacturer's instructions.

Techniques: Expressing, Ligation, Microscopy, RNA Extraction, Real-time Polymerase Chain Reaction, Control, Infection, Enzyme-linked Immunosorbent Assay

Journal: Infection and Immunity

Article Title: Porphyromonas gingivalis Exacerbates Ligature-Induced, RANKL-Dependent Alveolar Bone Resorption via Differential Regulation of Toll-Like Receptor 2 (TLR2) and TLR4

doi: 10.1128/IAI.02084-14

Figure Lengend Snippet: Alveolar bone resorption and gingival RANKL concentrations following 2 weeks of treatment in WT, Tlr2−/−, and Tlr4−/− mice. (A) Alveolar bone resorption around maxillary second molars (left and right), induced by ligation or ligation plus P. gingivalis infection, was analyzed after 2 weeks of treatment of WT, Tlr2−/−, and Tlr4−/− mice and is presented as the area of bone loss (mm2). (B) Total protein was isolated from gingival tissues, and the RANKL concentration was analyzed by ELISA using a specific antibody. The nontreatment groups were used as controls. Data are means and SE (n = 10).

Article Snippet: The RANKL levels in gingival homogenates were detected using a murine RANKL enzyme-linked immunosorbent assay (ELISA) development kit (Peprotech).

Techniques: Ligation, Infection, Isolation, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Infection and Immunity

Article Title: Porphyromonas gingivalis Exacerbates Ligature-Induced, RANKL-Dependent Alveolar Bone Resorption via Differential Regulation of Toll-Like Receptor 2 (TLR2) and TLR4

doi: 10.1128/IAI.02084-14

Figure Lengend Snippet: Bone resorption and gingival RANKL concentrations after ligature removal. Two weeks after treatment by ligation or ligation plus P. gingivalis infection, ligatures were removed from mice to allow wound healing for another 2 weeks. (A) The alveolar bone resorption around maxillary second molars (left and right) in WT, Tlr2−/−, and Tlr4−/− mice was measured and is presented as the area of bone loss (mm2). To further verify the observed difference in bone resorption between mice treated with ligation plus P. gingivalis and those treated with ligation alone, 3D bone assessment was performed using micro-CT to determine the remaining bone volume (mm3) (B), the bone volume density (% of bone surface/bone volume) (C), the trabecular thickness of remaining bone (mm) (D), and the trabecular number of remaining bone (1/mm) (E). (F) Total protein was isolated from gingival tissues, and RANKL production in gingival tissues was analyzed by ELISA using a specific antibody. Data are means and SE (n = 10). *, P < 0.05; **, P < 0.01.

Article Snippet: The RANKL levels in gingival homogenates were detected using a murine RANKL enzyme-linked immunosorbent assay (ELISA) development kit (Peprotech).

Techniques: Ligation, Infection, Micro-CT, Isolation, Enzyme-linked Immunosorbent Assay

Journal: Infection and Immunity

Article Title: Porphyromonas gingivalis Exacerbates Ligature-Induced, RANKL-Dependent Alveolar Bone Resorption via Differential Regulation of Toll-Like Receptor 2 (TLR2) and TLR4

doi: 10.1128/IAI.02084-14

Figure Lengend Snippet: Bone resorption and gingival RANKL and OPG expression after RANKL blockade in WT, Tlr2−/−, and Tlr4−/− mice. Two weeks after treatment by ligation or ligation plus P. gingivalis infection, ligatures were removed from mice to allow wound healing for another 2 weeks. During this 2-week period, mice were injected with either rabbit anti-mouse RANKL IgG or normal rabbit IgG in the interdental papilla (1 μg/site) on both the buccal and lingual sides of the maxillary second molars (left and right), on days 15, 17, and 21. (A) At the termination of the experiment (day 28), the alveolar bone resorption around maxillary second molars (left and right) in WT, Tlr2−/−, and Tlr4−/− mice was measured and is presented as the area of bone loss (mm2). (B) Total protein was isolated from gingival tissues, and gingival RANKL concentrations were measured by ELISA. (C) Total RNA was isolated from gingival tissues, and RANKL mRNA expression and OPG mRNA expression in gingival tissues were detected by real-time PCR. The RANKL/OPG ratio was then calculated. Amplification of the GAPDH gene was used as a control. L, ligation; P, P. gingivalis infection. Data are means and SE (n = 10). *, P < 0.05; **, P < 0.01.

Article Snippet: The RANKL levels in gingival homogenates were detected using a murine RANKL enzyme-linked immunosorbent assay (ELISA) development kit (Peprotech).

Techniques: Expressing, Ligation, Infection, Injection, Isolation, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Amplification