goat anti human cx3cl1 antibodies  (R&D Systems)

Journal: Journal of inflammation research

Article Title: Synergistic induction of CX3CL1 by TNF alpha and IFN gamma in osteoblasts from rheumatoid arthritis: involvement of NF-kappa B and STAT-1 signaling pathways

doi:

Figure Lengend Snippet: Secretion of CX3CL1 from OBs and the effects of TNF-α and IFN-γ. Cultured OBs from RA patients ( A ) or MG63 cells ( B ) were incubated for 24 h with the indicated doses of TNF-α and/or IFN-γ. ( C ) RA OBs stimulated with cytokines were collected after 2, 4, 8, and 24 h of stimulation. ( D ) RA OBs stimulated with cytokines were treated with either GM-6001 or negative control compound for 24 h. CX3CL1 levels in culture supernatants and cell lysates were assayed using specific ELISAs. Data are expressed as the means ± SEM of 5 independent experiments. * p < 0.01 vs control medium; ** p < 0.05 vs control medium.

Article Snippet: Monoclonal murine anti-human CX3CL1 (Genzyme/Techne, 4 μg/mL) was used as the primary antibody (Ab), and the secondary Ab was biotinylated polyclonal goat anti-CX3CL1 (Genzyme/Techne, 0.25 μg/mL).

Techniques: Cell Culture, Incubation, Negative Control

Journal: Journal of inflammation research

Article Title: Synergistic induction of CX3CL1 by TNF alpha and IFN gamma in osteoblasts from rheumatoid arthritis: involvement of NF-kappa B and STAT-1 signaling pathways

doi:

Figure Lengend Snippet: Cell surface expression and mRNA expression of CX3CL1 in RA OBs. ( A ) Representative photomicrographs showing the immunohistochemical localization of antigenic CX3CL1 in RA OBs stimulated with TNF-α (20 ng/mL) and IFN-γ (1000 U/mL) for 24 h (original magnification, 400×). ( B ) Flow cytometric analysis of the level of CXC3L1 expression. Cultured OBs from RA patients were incubated for 24 h with TNF-α (20 ng/mL) and IFN-γ (1000 U/mL). ( C ) Real-time RT-PCR analysis of CX3CL1 mRNA expression in RA OBs. Cultured OBs from RA patients were incubated for 4 h with TNF-α (20 ng/mL) and IFN-γ (1000 U/mL). Data are means ± SEM of 3 independent experiments. * p < 0.05 vs control medium. ( D ) Representative agarose gel analysis of CX3CL1 semiquantitative RT-PCR products.

Article Snippet: Monoclonal murine anti-human CX3CL1 (Genzyme/Techne, 4 μg/mL) was used as the primary antibody (Ab), and the secondary Ab was biotinylated polyclonal goat anti-CX3CL1 (Genzyme/Techne, 0.25 μg/mL).

Techniques: Expressing, Immunohistochemical staining, Cell Culture, Incubation, Quantitative RT-PCR, Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of inflammation research

Article Title: Synergistic induction of CX3CL1 by TNF alpha and IFN gamma in osteoblasts from rheumatoid arthritis: involvement of NF-kappa B and STAT-1 signaling pathways

doi:

Figure Lengend Snippet: Secretion ( A ) and mRNA expression ( B ) of CX3CL1 by OBs from RA, OA, or normal controls. OBs from RA patients (n = 3), OA patients (n = 2), or normal individuals (n = 2) were incubated for 24 h or 4 h with TNF-α (20 ng/mL) and IFN-γ (1000 U/mL). CX3CL1 levels in the culture supernatants ( A ) and CX3CL1 mRNA expression ( B ) were assayed using specific ELISAs and real-time PCR, respectively. Data are expressed as the means ± SEM of 3 to 5 independent experiments. * p < 0.05 vs control OBs, ** p < 0.05 vs control OBs and OA OBs.

Article Snippet: Monoclonal murine anti-human CX3CL1 (Genzyme/Techne, 4 μg/mL) was used as the primary antibody (Ab), and the secondary Ab was biotinylated polyclonal goat anti-CX3CL1 (Genzyme/Techne, 0.25 μg/mL).

Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction

Journal: Journal of inflammation research

Article Title: Synergistic induction of CX3CL1 by TNF alpha and IFN gamma in osteoblasts from rheumatoid arthritis: involvement of NF-kappa B and STAT-1 signaling pathways

doi:

Figure Lengend Snippet: NF-κB expression in OBs and its role in the induction of CX3CL1 expression. ( A ) Real-time RT-PCR analysis revealed enhanced expression of NF-κB mRNA in RA OBs incubated for 4 h with TNF-α (20 ng/mL) plus IFN-γ (1000 U/mL). ( B and C ) Effects of NF-κB-specific siRNA on CX3CL1 secretion and mRNA expression. RA OBs transfected with either NF-κB-specific siRNA or negative control siRNA were incubated for 24 h ( B ) or 4 h ( C ) with TNF-α (20 ng/mL) plus IFN-γ (1000 U/mL), after which culture supernatants ( B ) or total RNA ( C ) were isolated and ELISA or real-time PCR for CX3CL1 were conducted. Data are means ± SEM from 3 independent experiments. * p < 0.05 vs medium alone (A), * p < 0.05 vs TNF-α/IFN-γ with control siRNA ( B and C ).

Article Snippet: Monoclonal murine anti-human CX3CL1 (Genzyme/Techne, 4 μg/mL) was used as the primary antibody (Ab), and the secondary Ab was biotinylated polyclonal goat anti-CX3CL1 (Genzyme/Techne, 0.25 μg/mL).

Techniques: Expressing, Quantitative RT-PCR, Incubation, Transfection, Negative Control, Isolation, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Journal of inflammation research

Article Title: Synergistic induction of CX3CL1 by TNF alpha and IFN gamma in osteoblasts from rheumatoid arthritis: involvement of NF-kappa B and STAT-1 signaling pathways

doi:

Figure Lengend Snippet: Effect of pyrrolidine, an NF-κB inhibitor, on CX3CL1 expression by OBs. RA OBs stimulated with TNF-α/IFN-γ were incubated with graded doses of the NF-κB inhibitor pyrrolidine, after which culture supernatants ( A ) or total RNA ( B ) was isolated and ELISA or real-time PCR for CX3CL1 were conducted. Data are means ± SEM from three independent experiments. * p < 0.05 vs TNF-α/IFN-γ.

Article Snippet: Monoclonal murine anti-human CX3CL1 (Genzyme/Techne, 4 μg/mL) was used as the primary antibody (Ab), and the secondary Ab was biotinylated polyclonal goat anti-CX3CL1 (Genzyme/Techne, 0.25 μg/mL).

Techniques: Expressing, Incubation, Isolation, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Journal of inflammation research

Article Title: Synergistic induction of CX3CL1 by TNF alpha and IFN gamma in osteoblasts from rheumatoid arthritis: involvement of NF-kappa B and STAT-1 signaling pathways

doi:

Figure Lengend Snippet: STAT-1 expression in OBs and its role in the induction of CX3CL1 expression. ( A ) Real-time RT-PCR analysis revealed enhanced expression of STAT-1 mRNA in RA OBs incubated for 4 h with TNF-α (20 ng/mL) plus IFN-γ (1000 U/mL) and in RA OBs incubated with IFN-γ alone. ( B and C ) Effects of STAT-1-specific siRNA on CX3CL1 secretion and expression. RA OBs transfected with either STAT-1-specific siRNA or negative control siRNA were incubated for 24 h ( B ) or 4 h ( C ) with TNF-α (20 ng/mL) plus IFN-γ (1000 U/mL), after which culture supernatants ( B ) or total RNA ( C ) was isolated and ELISA or real-time PCR for CX3CL1 was conducted. Data are means ± SEM from three independent experiments. * p < 0.05 vs medium alone ( A ), * p < 0.05 vs TNF-α/IFN-γ with control siRNA ( B and C ).

Article Snippet: Monoclonal murine anti-human CX3CL1 (Genzyme/Techne, 4 μg/mL) was used as the primary antibody (Ab), and the secondary Ab was biotinylated polyclonal goat anti-CX3CL1 (Genzyme/Techne, 0.25 μg/mL).

Techniques: Expressing, Quantitative RT-PCR, Incubation, Transfection, Negative Control, Isolation, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction