rabbit anti capn1 antibody  (Proteintech)

Journal: International Dental Journal

Article Title: Calpain-1 Potentiates Periodontal Regeneration via PHLPP1-ERK-Driven Osteogenesis in Periodontal Ligament Stem Cells

doi: 10.1016/j.identj.2025.109309

Figure Lengend Snippet: CAPN1 contributed to the osteogenic differentiation process of hPDLSCs . (A) Oil Red O staining, ARS staining, as well as Alcian Blue staining. Scale bar = 100 μm. (B) Flow cytometry analysis for surface markers of hPDLSCs. (C) The Fluo-4 AM assay of hPDLSCs during osteogenic differentiation. Scale bar = 100 μm. (D) The expression of CAPN1 during osteogenic differentiation. (E) Calpain activity during osteogenic differentiation. Mean ± SD was employed to express all data (ns P > .05, ∗ P < .05, ∗∗ P < .01, *** P < .001, ∗∗∗∗ P < .0001).

Article Snippet: The 20 μg/mL antibody IgG (Santa Cruz) and CAPN1 (Santa Cruz) combined with the magnetic beads that had been treated, and the mixture was incubated overnight at 4°C in a flipping suspension apparatus (Bio-Rad).

Techniques: Staining, Flow Cytometry, Expressing, Activity Assay

Journal: International Dental Journal

Article Title: Calpain-1 Potentiates Periodontal Regeneration via PHLPP1-ERK-Driven Osteogenesis in Periodontal Ligament Stem Cells

doi: 10.1016/j.identj.2025.109309

Figure Lengend Snippet: CAPN1 promoted the osteogenic differentiation of hPDLSCs . (A) Representative images with CAPN1 -overexpressed in hPDLSCs. Scale bar = 50 μm. (B) Representative Western blot of CAPN1 in hPDLSCs after lentivirus transfection and quantification of the images. (C) Relative expression level of CAPN1 in hPDLSCs after lentivirus transfection. (D) ARS staining in hPDLSCs with or without lentivirus transfection. Scale bar = 200 μm. (E) Representative Western blot of CAPN1 expression in hPDLSCs following siRNAs treatment and quantification of the images. (F) Relative mRNA expression of CAPN1 in hPDLSCs following siRNAs treatment. (G) Relative mRNA expression of CAPN1, ALP, COL1Α1 , and RUNX2 on osteogenic day 0, 7, and 14 of hPDLSCs with or without lentivirus transfection. (H) ARS staining in hPDLSCs treated with siRNAs. Scale bar = 200 μm. (I) ARS staining in hPDLSCs following Calpeptin treatment. Scale bar = 200 μm. (J) Relative mRNA expression of CAPN1, ALP, COL1Α1 , and RUNX2 on osteogenic day 0,7, and 14 of hPDLSCs following siRNAs treatment. (K) Relative mRNA expression of CAPN1, ALP, COL1Α1 , and RUNX2 on osteogenic day 0, 7, and 14 of hPDLSCs following Calpeptin treatment. Mean ± SD was employed to express all data (ns P > .05, ∗ P < .05, ∗∗ P < .01, *** P < .001).

Article Snippet: The 20 μg/mL antibody IgG (Santa Cruz) and CAPN1 (Santa Cruz) combined with the magnetic beads that had been treated, and the mixture was incubated overnight at 4°C in a flipping suspension apparatus (Bio-Rad).

Techniques: Western Blot, Transfection, Expressing, Staining

Journal: International Dental Journal

Article Title: Calpain-1 Potentiates Periodontal Regeneration via PHLPP1-ERK-Driven Osteogenesis in Periodontal Ligament Stem Cells

doi: 10.1016/j.identj.2025.109309

Figure Lengend Snippet: CAPN1 facilitated the osteogenic differentiation of hPDLSCs through the ERK signalling pathway . (A) Representative Western blot for CAPN1, p-ERK1/2, and ERK1/2 in hPDLSCs with or without lentivirus transfection. (B) Quantification of the images in A. (C) Representative Western blot for CAPN1, p-ERK1/2, and ERK1/2 in hPDLSCs following siRNAs treatment. (D) Quantification of the images in C. (E) Representative Western blot for PHLPP1 in hPDLSCs following siRNAs treatment and quantification of the images. (F) Quantification of the images in E. (G) Representative Western blot for PHLPP1 in hPDLSCs following Calpeptin treatment. (H) Quantification of the images in G. (I) Representative Western blot for CAPN1, ERK1/2, and p-ERK1/2 in hPDLSCs following PD98059 or Calpeptin treatment. (J) Quantification of the images in J. (K) Representative Western blot for PHLPP1, ERK1/2, p-ERK1/2 in hPDLSCs following increasing NSC45586 doses treatment. (L) Quantification of the images in J. (M) Representative Western blot for PHLPP1, ERK1/2, p-ERK1/2 in hPDLSCs following siPHLPP1 treatment. (N) Quantification of the images in M. (O) Representative Western blot for PHLPP1, ERK1/2, p-ERK1/2 in hPDLSCs following si CAPN1 or si CAPN1 combined with NSC45586 treatment. (P) Quantification of the images in O. Mean ± SD was employed to express all data (ns P > .05, ∗ P < .05, ∗∗ P < .01, *** P < .001, **** P < .0001).

Article Snippet: The 20 μg/mL antibody IgG (Santa Cruz) and CAPN1 (Santa Cruz) combined with the magnetic beads that had been treated, and the mixture was incubated overnight at 4°C in a flipping suspension apparatus (Bio-Rad).

Techniques: Western Blot, Transfection

Journal: International Dental Journal

Article Title: Calpain-1 Potentiates Periodontal Regeneration via PHLPP1-ERK-Driven Osteogenesis in Periodontal Ligament Stem Cells

doi: 10.1016/j.identj.2025.109309

Figure Lengend Snippet: CAPN1 binds to PHLPP1 during osteogenic differentiation of h PDLSCs . (A) Representative immunofluorescence images of CAPN1 and PHLPP1 on 0, 3, 6, 9, 12, 15, 18, and 21 days in hPDLSCs. Green: CAPN1; Red: PHLPP1; Blue: DAPI. Scale bar = 200 μm. (B) Intensity profiles showing signals from two fluorescent channels. (C) CAPN1 and PHLPP1 molecular docking structure. CAPN1 and PHLPP1 are depicted in green and blue, respectively. Hydrogen bonds are indicated in yellow. (D-H) The results of molecular dynamics (MD) simulations. (D) Root mean square deviation (RMSD) of the complex. (E) Root mean square fluctuation (RMSF) of the complex. (F) Radius of gyration (Rg) change of the complex. (G) The number of hydrogen bonds of the complex. (H) Solvent-accessible surface area (SASA) change of the complex. (I) Co-IP assay showing representative protein bands of CAPN1 and PHLPP1 in HEK293T.

Article Snippet: The 20 μg/mL antibody IgG (Santa Cruz) and CAPN1 (Santa Cruz) combined with the magnetic beads that had been treated, and the mixture was incubated overnight at 4°C in a flipping suspension apparatus (Bio-Rad).

Techniques: Immunofluorescence, Solvent, Co-Immunoprecipitation Assay

Journal: International Dental Journal

Article Title: Calpain-1 Potentiates Periodontal Regeneration via PHLPP1-ERK-Driven Osteogenesis in Periodontal Ligament Stem Cells

doi: 10.1016/j.identj.2025.109309

Figure Lengend Snippet: CAPN1 promoted ectopic bone regeneration. (A and B) Representative pictures of H&E and Masson staining of new bone-like tissue formation in HA/TCP constructs. Scale bar = 200 μm for the upper row of A and B. Scale bar = 50 μm for the lower row of A and B. Yellow arrows indicate new bone-like tissue. Magnified images (yellow squares) were captured at the area of new bone-like tissue formation. (C) Immunofluorescence staining was used to visualize the expression of OPN, PHLPP1, and p-ERK1/2 in HA/TCP constructs. Red: OPN, PHLPP1, p-ERK1/2; Blue: DAPI. Scale bars = 50 μm.

Article Snippet: The 20 μg/mL antibody IgG (Santa Cruz) and CAPN1 (Santa Cruz) combined with the magnetic beads that had been treated, and the mixture was incubated overnight at 4°C in a flipping suspension apparatus (Bio-Rad).

Techniques: Staining, Construct, Immunofluorescence, Expressing

Journal: International Dental Journal

Article Title: Calpain-1 Potentiates Periodontal Regeneration via PHLPP1-ERK-Driven Osteogenesis in Periodontal Ligament Stem Cells

doi: 10.1016/j.identj.2025.109309

Figure Lengend Snippet: CAPN1 promoted the periodontal bone repair and regeneration . (A) 3D reconstruction of micro-CT. (B) Representative images of micro-CT. Yellow dotted line represents the site of the defect. Scale bar = 1 mm. (C) Micro-CT analysis of newly formed bone in different time across various groups. (D-E) Representative pictures of H&E and Masson staining of the area of new bone formation in different time. Magnified images (yellow squares) were captured at the centre and edges of the defect. Scale bar = 200 μm for the upper row, Scale bar = 100 μm for the lower row. Mean ± SD was employed to express all data (ns P > .05, ∗ P < .05, ∗∗ P < .01, *** P < .001).

Article Snippet: The 20 μg/mL antibody IgG (Santa Cruz) and CAPN1 (Santa Cruz) combined with the magnetic beads that had been treated, and the mixture was incubated overnight at 4°C in a flipping suspension apparatus (Bio-Rad).

Techniques: Micro-CT, Staining

Journal: International Dental Journal

Article Title: Calpain-1 Potentiates Periodontal Regeneration via PHLPP1-ERK-Driven Osteogenesis in Periodontal Ligament Stem Cells

doi: 10.1016/j.identj.2025.109309

Figure Lengend Snippet: Graphical illustration of CAPN1 potentiates osteogenic differentiation of hPDLSCs via the PHLPP1/ERK1/2 axis . hPDLSCs were isolated from impacted third molars were extracted from patients. hPDLSCs were loaded into GelMa hydrogel and implanted into mandibular periodontal bone defects. During the osteogenic differentiation process, extracellular Ca²⁺ enters the cell to activate CAPN1. Activated CAPN1 induces degradation of PHLPP1, which in turn promotes phosphorylation of ERK1/2. Phosphorylated ERK1/2 translocates into the nucleus and regulates the expression of osteogenesis-related genes.

Article Snippet: The 20 μg/mL antibody IgG (Santa Cruz) and CAPN1 (Santa Cruz) combined with the magnetic beads that had been treated, and the mixture was incubated overnight at 4°C in a flipping suspension apparatus (Bio-Rad).

Techniques: Isolation, Phospho-proteomics, Expressing