anti rat nod1  (Novus Biologicals)

Journal: BioMed Research International

Article Title: Functional Roles of NOD1 in Odontoblasts on Dental Pulp Innate Immunity

doi: 10.1155/2016/9325436

Figure Lengend Snippet: Constitutive expression of NOD1 and NOD2 in rat KN-3 cells and chemokine production in iE-DAP- and TNF- α -stimulated KN-3 cells. (a) The constitutive expression of NOD1 and NOD2 in KN-3 cells was assessed by flow cytometry. The results are representative of four different experiments demonstrating similar results. (b, c) KN-3 cells were stimulated with iE-DAP (10 μ g mL −1 ), iE-Lys (10 μ g mL −1 ), MDP (10 μ g mL −1 ), or TNF- α (0.01 μ g mL −1 ) for 24 h. The detection of cytokines and chemokines produced in the cell culture supernatants was performed using a Rat Cytokine Antibody Array. (b) The results shown are representative images of two independent experiments with similar results. (c) Densitometric analysis of various chemokine production levels. Bars indicate the relative densitometric intensities after the values were normalized with both positive and negative controls and background using ImageJ software. In particular, positive controls were used to normalize the values from different membranes being compared. Values represent the means ± SDs of two independent experiments. Asterisks indicate significant differences ( ∗ p < 0.05 and ∗∗ p < 0.01) versus nonstimulated control group.

Article Snippet: KN-3 cells fixed in 4% paraformaldehyde were treated with BD Cytofix/Cytoperm solution (BD Biosciences, San Jose, CA, USA), incubated with anti-rat NOD1 (NOVUS BIOLOGICALS, Littleton, CO, USA), NOD2 (Abnova, Jhouzih St., Taipei, Taiwan) antibodies or their respective isotype-matched controls followed by reaction with fluorescein isothiocyanate- (FITC-) conjugated rabbit IgG secondary antibody (Dako, Carpinteria, CA, USA).

Techniques: Expressing, Flow Cytometry, Produced, Cell Culture, Ab Array, Software, Control

Journal: BioMed Research International

Article Title: Functional Roles of NOD1 in Odontoblasts on Dental Pulp Innate Immunity

doi: 10.1155/2016/9325436

Figure Lengend Snippet: Expression and production of chemokines induced in KN-3 cells stimulated with NOD1 or NOD2 ligand. (a, b) KN-3 cells were stimulated with iE-DAP (1 or 10 μ g mL −1 ), iE-Lys (10 μ g mL −1 ), MDP (1 or 10 μ g mL −1 ), MDP-LL (10 μ g mL −1 ), or TNF- α (0.01 μ g mL −1 ) for 24 h. (a) After stimulation, total RNA was isolated and mRNA expression levels of CINC-1, CINC-2 α , MCP-1, and CCL20 were analyzed by real-time reverse transcription-polymerase chain reaction (RT-PCR). (b) The concentrations of CINC-1, CINC-2, MCP-1, and CCL20 in the cell culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA). (c) KN-3 cells were stimulated with iE-DAP (10 μ g mL −1 ), iE-Lys (10 μ g mL −1 ), or TNF- α (0.01 μ g mL −1 ) for 4, 12, or 24 h. After stimulation, total RNA was isolated and mRNA expression levels of CINC-1, CINC-2 α , MCP-1, and CCL20 were analyzed by real-time RT-PCR. Values represent the means ± SDs from representative of four independent experiments and each experiment was performed in quadruplicate. Asterisks indicate significant differences ( ∗ p < 0.05 and ∗∗ p < 0.01) versus nonstimulated control group.

Article Snippet: KN-3 cells fixed in 4% paraformaldehyde were treated with BD Cytofix/Cytoperm solution (BD Biosciences, San Jose, CA, USA), incubated with anti-rat NOD1 (NOVUS BIOLOGICALS, Littleton, CO, USA), NOD2 (Abnova, Jhouzih St., Taipei, Taiwan) antibodies or their respective isotype-matched controls followed by reaction with fluorescein isothiocyanate- (FITC-) conjugated rabbit IgG secondary antibody (Dako, Carpinteria, CA, USA).

Techniques: Expressing, Isolation, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Control