rabbit anti dmp 1 polyclonal antibody  (TaKaRa)

Journal: Molecules

Article Title: RORα Regulates Odontoblastic Differentiation and Mediates the Pro-Odontogenic Effect of Melatonin on Dental Papilla Cells

doi: 10.3390/molecules26041098

Figure Lengend Snippet: Figure 1. Expression of nuclear receptor retinoid acid receptor-related orphan receptors (RORs) in rat dental papilla cells (rDPCs). (a) Total RNA was extracted from rDPCs and detected by reverse transcription polymerase chain reaction (RT-PCR) and agarose gel electrophoresis assay using specific primers for RORα, RORβ, RORγ, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). RT-PCR without primers served as the negative control (NC); (b) Immunofluorescence staining was performed with polyclonal anti-RORα antibody (green), and nuclei were labelled with 4-6-diamidino-2-phenylindole (DAPI, blue). Scale bar: 50 µm.

Article Snippet: Subsequently, they were incubated overnight at 4 ◦C with the following primary antibodies: polyclonal rabbit anti-RORα antibody (1:2000, PA5-23268, Thermo Scientific, MA, USA), monoclonal mouse anti-DSPP antibody (1:500, sc-73632, Santa Cruz, CA, USA), polyclonal rabbit anti-DMP1 antibody (1:1000, NBP 1-45525, Novus Biologicals, Littleton, CO, USA), and monoclonal mouse anti-β-actin (1:1000, AF0003, Beyotime, Shanghai, China).

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Negative Control, Staining

Journal: Molecules

Article Title: RORα Regulates Odontoblastic Differentiation and Mediates the Pro-Odontogenic Effect of Melatonin on Dental Papilla Cells

doi: 10.3390/molecules26041098

Figure Lengend Snippet: Figure 2. The expression pattern of retinoid acid receptor-related orphan receptor α (RORα) and odontogenic markers in rat dental papilla cells (rDPCs) during odontoblastic differentiation. rDPCs were cultured in maintained medium (control) or odontogenic induction medium (OS) for 3 and 7 days (3d and 7d), respectively. (a) The mRNA levels of RORα, dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP1), and alkaline phosphatase (ALP) were quantified by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Glyceraldehyde-3-phosphate dehydroge- nase (GAPDH) was used as the normalisation control; (b) The protein levels of RORα, DSPP, and DMP1 were determined by western blotting and normalised to the protein level of β-actin. All data are presented as the mean ± SD (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Control group.

Article Snippet: Subsequently, they were incubated overnight at 4 ◦C with the following primary antibodies: polyclonal rabbit anti-RORα antibody (1:2000, PA5-23268, Thermo Scientific, MA, USA), monoclonal mouse anti-DSPP antibody (1:500, sc-73632, Santa Cruz, CA, USA), polyclonal rabbit anti-DMP1 antibody (1:1000, NBP 1-45525, Novus Biologicals, Littleton, CO, USA), and monoclonal mouse anti-β-actin (1:1000, AF0003, Beyotime, Shanghai, China).

Techniques: Expressing, Cell Culture, Control, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot

Journal: Molecules

Article Title: RORα Regulates Odontoblastic Differentiation and Mediates the Pro-Odontogenic Effect of Melatonin on Dental Papilla Cells

doi: 10.3390/molecules26041098

Figure Lengend Snippet: Figure 4. Effect of retinoid acid receptor-related orphan receptor α (RORα) overexpression on odontoblastic differentiation of rat dental papilla cells (rDPCs). rDPCs were transfected with pcDNA3.1-RORα (RORα group) or pcDNA3.1-NC (negative control group) for 24 h and then cultured in control or odontogenic induction (OS) medium for 3 or 7 days. (a) The transfection efficiency of RORα overexpression was assessed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting. (b) The mRNA levels of dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP1), and alkaline phosphatase (ALP) were detected by qRT-PCR after 3-day induction. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the normalisation control; (c) The protein levels of DSPP and DMP1 were measured by western blotting after 7-day induction. β-actin was used as the internal control; (d) ALP activity in cellular lysates was determined after 7-day odontogenic induction. (e) The formation of mineralized nodules was visualized by alizarin red staining at 7 days after induction. Scale bar: 100 µm. All data are presented as the mean ± SD (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. pcDNA3.1-NC or Control-NC group, # p < 0.05, ### p < 0.001 vs. OS-NC group.

Article Snippet: Subsequently, they were incubated overnight at 4 ◦C with the following primary antibodies: polyclonal rabbit anti-RORα antibody (1:2000, PA5-23268, Thermo Scientific, MA, USA), monoclonal mouse anti-DSPP antibody (1:500, sc-73632, Santa Cruz, CA, USA), polyclonal rabbit anti-DMP1 antibody (1:1000, NBP 1-45525, Novus Biologicals, Littleton, CO, USA), and monoclonal mouse anti-β-actin (1:1000, AF0003, Beyotime, Shanghai, China).

Techniques: Over Expression, Transfection, Negative Control, Cell Culture, Control, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Activity Assay, Staining

Journal: Molecules

Article Title: RORα Regulates Odontoblastic Differentiation and Mediates the Pro-Odontogenic Effect of Melatonin on Dental Papilla Cells

doi: 10.3390/molecules26041098

Figure Lengend Snippet: Figure 5. Effect of retinoid acid receptor-related orphan receptor α (RORα) knockdown on odontoblastic differentiation of rat dental papilla cells (rDPCs). rDPCs were transfected with small interfering (si) RORα or si negative control (NC) for 8 h and then cultured in control or odontogenic induction (OS) medium for 3 or 7 days. (a) The knockdown efficiency of RORα was examined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting; (b) The mRNA levels of dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP1), and alkaline phosphatase (ALP) were measured by qRT-PCR after 3-day induction. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the internal control; (c) The protein levels of DSPP and DMP1 were detected by western blotting after 7-day induction and normalised to β-actin levels; (d) ALP activity was analysed after 7 days of odontogenic induction. (e) The formation of mineralized nodules was assessed by alizarin red staining at day 7. Scale bar: 100 µm. All data are presented as the mean ± SD (n = 3). ** p < 0.01, *** p < 0.001 vs. si NC or Control-si NC group, # p < 0.05, ## p < 0.01 vs. OS-si NC group.

Article Snippet: Subsequently, they were incubated overnight at 4 ◦C with the following primary antibodies: polyclonal rabbit anti-RORα antibody (1:2000, PA5-23268, Thermo Scientific, MA, USA), monoclonal mouse anti-DSPP antibody (1:500, sc-73632, Santa Cruz, CA, USA), polyclonal rabbit anti-DMP1 antibody (1:1000, NBP 1-45525, Novus Biologicals, Littleton, CO, USA), and monoclonal mouse anti-β-actin (1:1000, AF0003, Beyotime, Shanghai, China).

Techniques: Knockdown, Transfection, Negative Control, Cell Culture, Control, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Activity Assay, Staining

Journal: Contemporary Clinical Dentistry

Article Title: Clinical, Histological, and Molecular Perspective on Regenerating Nonvital Immature Teeth

doi: 10.4103/ccd.ccd_44_23

Figure Lengend Snippet: Immunohistochemical analysis. Dentin matrix protein-1 (DMP-1) and dentin sialoprotein (DSP) localization in newly formed tissue. Staining of respective positive technique controls for (a) DMP-1 (mammary gland) and (g) DSP (placenta) and negative technique control in the tooth (b and h). The positive control in the tooth for DMP-1 (c) and DSP (i) shows a layer of columnar odontoblasts surrounding the inner surface of dentin and positive staining for both odontogenic proteins as observed for the revascularized tooth (d and j). The intense staining of DMP-1 and DSP on the dentin mineralization front in the newly formed tissue (f and l) is similar to the controls (e and k). Bar corresponds to 100 μm

Article Snippet: The sections were incubated with primary antibodies overnight at 4°C: (1) rabbit polyclonal anti-DSPP (Abcam, Cambridge, MA, USA), which is specific for the N-DSPP, corresponding to the natural cleavage fragment of DSP, and (2) rabbit polyclonal anti-DMP-1 (Sigma-Aldrich, St Louis, MO, USA).

Techniques: Immunohistochemical staining, Staining, Positive Control