rabbit polyclonal anti type ii collagen  (Bioss)

Journal: Bioengineering

Article Title: Comparison of Bioengineered Scaffolds for the Induction of Osteochondrogenic Differentiation of Human Adipose-Derived Stem Cells

doi: 10.3390/bioengineering11090920

Figure Lengend Snippet: hASC survival and differentiation in GelMA hydrogels. ( A ) CCK8 assay of hASCs encapsulated in the GelMA hydrogel at different time points (1 week intervals). Results are reported as mean ± SEM of n = 3 samples/group. ( B ) Representative confocal imaging of Dead/live fluorescence staining of hASCs in the GelMA hydrogel after 3 weeks of culture. Red cells are dead while green cells are alive. ( C ) Alcian blue staining of GelMA hydrogel after 24 h and 3 weeks of culture. ( D ) Alizarin red staining of GelMA hydrogel after 24 h and 3 weeks of culture. ( E , F ) Confocal imaging of immunofluorescence for COL2A1 (green) and OCN (red) in hASCs encapsulated in GelMA hydrogel after 3 weeks of culture.

Article Snippet: Briefly, samples were incubated with rabbit polyclonal anti-collagen type II alpha 1 (COL2A1) antibody (Merck) and rabbit polyclonal anti-osteocalcin (OCN) antibody (Thermo Fisher Scientific) for 1 h at room temperature.

Techniques: CCK-8 Assay, Imaging, Fluorescence, Staining, Immunofluorescence

Journal: Bioengineering

Article Title: Comparison of Bioengineered Scaffolds for the Induction of Osteochondrogenic Differentiation of Human Adipose-Derived Stem Cells

doi: 10.3390/bioengineering11090920

Figure Lengend Snippet: hASC survival and differentiation in PEGDA scaffold. ( A ) CCK8 assay of hASCs seeded in the PEGDA scaffold at different time points (1 week intervals). Results are reported as mean ± SEM of n = 3 samples/group. T test: * p < 0.05. ( B ) Representative confocal imaging of Dead/live fluorescence staining of hASCs seeded in the PEGDA scaffold after 3 weeks of culture. ( C ) Confocal imaging of immunofluorescence for COL2A1 (green) and OCN (red) in hASCs seeded in the PEGDA scaffold after 3 weeks of culture.

Article Snippet: Briefly, samples were incubated with rabbit polyclonal anti-collagen type II alpha 1 (COL2A1) antibody (Merck) and rabbit polyclonal anti-osteocalcin (OCN) antibody (Thermo Fisher Scientific) for 1 h at room temperature.

Techniques: CCK-8 Assay, Imaging, Fluorescence, Staining, Immunofluorescence

Journal: Bioengineering

Article Title: Comparison of Bioengineered Scaffolds for the Induction of Osteochondrogenic Differentiation of Human Adipose-Derived Stem Cells

doi: 10.3390/bioengineering11090920

Figure Lengend Snippet: hASC survival and differentiation in celery-based scaffold. ( A ) SEM imaging of a single hASC inside a niche of the celery-based scaffold. ( B ) CCK8 assay of hASCs seeded in the celery-based scaffold at different time points (1 week intervals). Results are reported as mean ± SEM of n = 3 samples/group. T test: * p < 0.05. ( C ) Representative confocal imaging of Dead/live fluorescence staining of hASCs seeded in the scaffold after 3 weeks of culture. ( D ) Confocal 3D stack and ( E ) the projection of hASC distribution inside the scaffold. ( F ) Confocal imaging of immunofluorescence for COL2A1 (green) and OCN (red) in hASCs seeded in the PEGDA scaffold after 3 weeks of culture.

Article Snippet: Briefly, samples were incubated with rabbit polyclonal anti-collagen type II alpha 1 (COL2A1) antibody (Merck) and rabbit polyclonal anti-osteocalcin (OCN) antibody (Thermo Fisher Scientific) for 1 h at room temperature.

Techniques: Imaging, CCK-8 Assay, Fluorescence, Staining, Immunofluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: IRF1 regulation of ZBP1 links mitochondrial DNA and chondrocyte damage in osteoarthritis

doi: 10.1186/s12964-024-01744-1

Figure Lengend Snippet: ZBP1 is essential for chondrocyte damage. Chondrocytes were transfected with siNC or ZBP1 siRNA for 24 h following TNF-α induction for 24 h. (A, B) Western blots and quantitative analysis of ZBP1, MMP13, and MMP3 expression levels ( n = 3). (C, D) Western blots and quantitative analysis of SOX9 expression levels. (E, F) Western blot and quantitative analysis of iNOS expression levels ( n = 3). (G) qPCR results showing the relative expression levels of Zbp1 and Inos ( n = 3). (H-K) Immunofluorescence staining and fluorescence intensity analysis of the relative expression levels of COL2A1 and MMP13 ( n = 3; scale bar: 50 μm). Chondrocytes were transfected with siNC or ZBP1 siRNA following TSZ (20 ng/ml TNF-α, 100 nM Smac mimetic, and 20 mM Z-VAD) induction for 12 h. (L, M) Western blots and quantitative analysis of ZBP1, P-RIPK3, and P-MLKL expression levels ( n = 3). The data are shown as the means ± SDs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: COL2A1 Rabbit Polyclonal antibody (28459-1-AP; Western blot: 1:1000; IF: 1:200; IHC: 1:800), β-ACTIN Recombinant Rabbit antibody (81115-1-RR; Western blot: 1:10000), Phospho-RIPK1 (Ser161) Mouse Monoclonal antibody (66854-1-Ig; Western blot: 1:5000; IF: 1:400; IHC: 1:500), IRF1 Rabbit Polyclonal antibody (11335-1-AP; Western blot: 1:500; IF: 1:250), GAPDH Mouse mAb (60004-1-Ig; Western blot: 1:10000) and MMP13 Rabbit Polyclonal antibody (18165-1-AP; Western blot: 1:1000; IF: 1:200; IHC: 1:100) were acquired from Proteintech Group (Wuhan, China).

Techniques: Transfection, Western Blot, Expressing, Immunofluorescence, Staining, Fluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: IRF1 regulation of ZBP1 links mitochondrial DNA and chondrocyte damage in osteoarthritis

doi: 10.1186/s12964-024-01744-1

Figure Lengend Snippet: ZBP1 is essential for OA progression in DMM mice. (A, B) IHC staining showing ZBP1-positive cells in the cartilage of the four groups (scale bar: 100 μm). (C, D) Safranin O/fast green staining, H&E staining and OARSI scores of the four groups (sham + AAV9-shGFP, n = 8; sham + AAV9-shZBP1, n = 8; DMM + AAV9-shGFP, n = 8; DMM + AAV9-shZBP1, n = 8; scale bar: 200 μm). (E-H) IHC staining showing COL2A1-positive cells and MMP13-positive cells in the cartilage of the four groups (scale bar: 100 μm). (I) Images showing 3D reconstructions of mouse joints; the black arrow indicates an osteophyte (scale bar: 1 mm). (J, K) Transverse sectional images and statistical analysis of the maximum osteophyte outgrowth of each joint in the four groups; the white arrow indicates the osteophyte (scale bar: 1 mm). The data are shown as the means ± SDs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: COL2A1 Rabbit Polyclonal antibody (28459-1-AP; Western blot: 1:1000; IF: 1:200; IHC: 1:800), β-ACTIN Recombinant Rabbit antibody (81115-1-RR; Western blot: 1:10000), Phospho-RIPK1 (Ser161) Mouse Monoclonal antibody (66854-1-Ig; Western blot: 1:5000; IF: 1:400; IHC: 1:500), IRF1 Rabbit Polyclonal antibody (11335-1-AP; Western blot: 1:500; IF: 1:250), GAPDH Mouse mAb (60004-1-Ig; Western blot: 1:10000) and MMP13 Rabbit Polyclonal antibody (18165-1-AP; Western blot: 1:1000; IF: 1:200; IHC: 1:100) were acquired from Proteintech Group (Wuhan, China).

Techniques: Immunohistochemistry, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: IRF1 regulation of ZBP1 links mitochondrial DNA and chondrocyte damage in osteoarthritis

doi: 10.1186/s12964-024-01744-1

Figure Lengend Snippet: ZBP1 overexpression depends on IRF1. Chondrocytes were transfected with siNC or IRF1 siRNA for 24 h following TNF-α induction for 12 h. (A) Prediction of transcription factors using the UCSC Genome Browser database, SPP-ominer database, and Cistrome Data Browser. (B) qPCR results showing the relative mRNA expression levels of Irf1 and Zbp1 ( n = 3). (C, D) Western blots and quantitative analysis of the relative protein expression level of IRF1 ( n = 3). (E, F) Immunofluorescence staining and fluorescence intensity analysis of ZBP1 expression in chondrocytes transfected with siNC or IRF1 siRNA following TNF-α induction for 12 h ( n = 3; scale bar: 50 μm). (G-L) Western blots and quantitative analysis of the relative protein expression levels of iNOS, COX2, MMP3, MMP13, AGGRECAN, COL2A1, and SOX9 ( n = 3). (M-P) Immunofluorescence staining and fluorescence intensity analysis of COL2A1 and MMP13 expression in chondrocytes transfected with siNC or IRF1 siRNA following TNF-α induction for 12 h ( n = 3; scale bar: 50 μm). Chondrocytes were transfected with negative control siRNA(siNC) or IRF1 siRNA (si- Irf1 ) or ZBP1-plasmid (OE- Zbp1 ) or empty plasmid (OE-NC) for 24 h following TNF-α induction for 24 h. (Q-V) Western blots and quantitative analysis of the relative protein expression of AGGRECAN, COL2A1, SOX9, iNOS, COX2 MMP13, and MMP3 in chondrocytes ( n = 3). The data are shown as the means ± SDs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: COL2A1 Rabbit Polyclonal antibody (28459-1-AP; Western blot: 1:1000; IF: 1:200; IHC: 1:800), β-ACTIN Recombinant Rabbit antibody (81115-1-RR; Western blot: 1:10000), Phospho-RIPK1 (Ser161) Mouse Monoclonal antibody (66854-1-Ig; Western blot: 1:5000; IF: 1:400; IHC: 1:500), IRF1 Rabbit Polyclonal antibody (11335-1-AP; Western blot: 1:500; IF: 1:250), GAPDH Mouse mAb (60004-1-Ig; Western blot: 1:10000) and MMP13 Rabbit Polyclonal antibody (18165-1-AP; Western blot: 1:1000; IF: 1:200; IHC: 1:100) were acquired from Proteintech Group (Wuhan, China).

Techniques: Over Expression, Transfection, Expressing, Western Blot, Immunofluorescence, Staining, Fluorescence, Negative Control, Plasmid Preparation

Journal: Cell Communication and Signaling : CCS

Article Title: IRF1 regulation of ZBP1 links mitochondrial DNA and chondrocyte damage in osteoarthritis

doi: 10.1186/s12964-024-01744-1

Figure Lengend Snippet: Inhibition of the mtDNA-IRF1-ZBP1 axis with CsA protects mice from DMM-induced OA progression. (A, B) Safranin O/fast green staining, HE staining and OARSI scores of the three groups (DMM, n = 10; DMM + CsA (2 µg/kg), n = 10; DMM + CsA (20 µg/kg), n = 10) (scale bar: 200 μm). (C, D) IHC staining showing p-P65-positive cells and p-RIPK1-positive cells in the cartilage of the three groups (scale bar: 100 μm). (E, F) IHC staining showing COL2A1-positive cells and MMP13-positive cells in the cartilage of the three groups (scale bar: 100 μm). (G) Images of 3D reconstructions of mouse joints; the black arrow indicates the osteophyte (scale bar: 1 mm). (H, I) Transverse sectional images and statistical analysis of the maximum osteophyte outgrowth of each joint in the three groups; the white arrow indicates the osteophyte (DMM, n = 8; DMM + CsA (2 µg/kg), n = 8; DMM + CsA (20 µg/kg), n = 8; scale bar: 1 mm). The data are shown as the means ± SDs. * P < 0.05, ** P < 0.01, NS indicates not significant

Article Snippet: COL2A1 Rabbit Polyclonal antibody (28459-1-AP; Western blot: 1:1000; IF: 1:200; IHC: 1:800), β-ACTIN Recombinant Rabbit antibody (81115-1-RR; Western blot: 1:10000), Phospho-RIPK1 (Ser161) Mouse Monoclonal antibody (66854-1-Ig; Western blot: 1:5000; IF: 1:400; IHC: 1:500), IRF1 Rabbit Polyclonal antibody (11335-1-AP; Western blot: 1:500; IF: 1:250), GAPDH Mouse mAb (60004-1-Ig; Western blot: 1:10000) and MMP13 Rabbit Polyclonal antibody (18165-1-AP; Western blot: 1:1000; IF: 1:200; IHC: 1:100) were acquired from Proteintech Group (Wuhan, China).

Techniques: Inhibition, Staining, Immunohistochemistry