Journal: Experimental and Therapeutic Medicine

Article Title: Biotin-avidin mediates the binding of adipose-derived stem cells to a porous β-tricalcium phosphate scaffold: Mandibular regeneration

doi: 10.3892/etm.2015.2961

Figure Lengend Snippet: Comparison of the morphology of Bio-ADSCs on Avi-β-TCP and untreated ADSCs on β-TCP. Scanning electron microscopy micrographs of (A) ADSCs/β-TCP and (B) Bio-ADSCs/Avi-β-TCP constructs following 7 day incubation (magnification, ×1,200). ADSCs, adipose-derived stem cells; β-TCP, β-tricalcium phosphate; Bio-ADSCs, biotinylated ADSCs; Avi-β-TCP, avidin-coated β-TCP.

Article Snippet: After rinsing in PBS three times, the tissue sections were incubated with biotinylated secondary antibody (1:500; Z0420; DAKO, Glostrup, Denmark) at room temperature for 1 h. Peroxidase activity was visualized using 0.05% diaminobenzidine and 0.03% H2O2 in PBS.

Techniques: Electron Microscopy, Construct, Incubation, Derivative Assay, Avidin-Biotin Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Biotin-avidin mediates the binding of adipose-derived stem cells to a porous β-tricalcium phosphate scaffold: Mandibular regeneration

doi: 10.3892/etm.2015.2961

Figure Lengend Snippet: (A and B) Hematoxylin and eosin staining and (C and D) immunohistochemical analysis of osteocalcin protein expression at the mandibular defect area in the (A and C) group 1 and (B and D) group 2 rabbits at 4 weeks post-operation (magnification, ×200). The group 1 and group 2 rabbits received the Bio-ADSCs/Avi-β-TCP and Bio-ADSCs/Avi-β-TCP/PRP constructs, respectively. ADSCs, adipose-derived stem cells; β-TCP, β-tricalcium phosphate; Bio-ADSCs, biotinylated-ADSCs; Avi-β-TCP, avidin-coated β-TCP; PRP, platelet-rich plasma.

Article Snippet: After rinsing in PBS three times, the tissue sections were incubated with biotinylated secondary antibody (1:500; Z0420; DAKO, Glostrup, Denmark) at room temperature for 1 h. Peroxidase activity was visualized using 0.05% diaminobenzidine and 0.03% H2O2 in PBS.

Techniques: Staining, Immunohistochemistry, Expressing, Construct, Derivative Assay, Avidin-Biotin Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Biotin-avidin mediates the binding of adipose-derived stem cells to a porous β-tricalcium phosphate scaffold: Mandibular regeneration

doi: 10.3892/etm.2015.2961

Figure Lengend Snippet: Three-dimensional computed tomography images of mandibular defect reconstruction in the (A) group 1 and (B) group 2 rabbits at 4 weeks post-operation (red arrow). The group 1 and group 2 rabbits received the Bio-ADSCs/Avi-β-TCP and Bio-ADSCs/Avi-β-TCP/PRP constructs, respectively. ADSCs, adipose-derived stem cells; β-TCP, β-tricalcium phosphate; Bio-ADSCs, biotinylated-ADSCs; Avi-β-TCP, avidin-coated β-TCP; PRP, platelet-rich plasma.

Article Snippet: After rinsing in PBS three times, the tissue sections were incubated with biotinylated secondary antibody (1:500; Z0420; DAKO, Glostrup, Denmark) at room temperature for 1 h. Peroxidase activity was visualized using 0.05% diaminobenzidine and 0.03% H2O2 in PBS.

Techniques: Computed Tomography, Construct, Derivative Assay, Avidin-Biotin Assay

Journal: Scientific Reports

Article Title: Dual role of the colonization factor CD2831 in Clostridium difficile pathogenesis

doi: 10.1038/s41598-019-42000-8

Figure Lengend Snippet: Binding of recombinant CD2831 full-length protein and subdomains to collagen. ( A ) Schematic representation of CD2831 domains organization displays the Leader Peptide at the N-terminus (orange) followed by a Short Repeat Region (yellow) in which repeated amino acids sequences are marked in bold, two collagen-binding domains (blue) and a Proline-rich region (green) recognized by the protease ZmpI/PPEP-1 that cleaves within the seven NPP amino acids sequences of the domain. The charged tail and the hydrophobic domain are present at the C-terminus (red). The protein is cleaved between the threonyl and glycil residues of the PPKTG motif and transferred to the cell-wall peptidoglycan by the Sortase B enzyme. Blue, black and grey lines indicate the length of recombinant CD2831 subdomains compared to the full-length. ( B ) Immunofluorescence of 1 µM of CD2831 (green) binding to collagen (red) produced by IMR90 cells. Collagen staining results from a mix of antibodies against collagen types I, III and V in a 1:1:1 ratio. Binding of the proteins was detected by using rabbit anti-CD2831 serum and secondary Alexa Fluor-conjugated antibodies. ( C ) Human fibroblasts IMR90 were cultured into collagen I-coated 96 well plates for three days after seeding. Cells were then incubated with serially diluted recombinant CD2831 proteins ranging from 0.08 to 10 µM. Binding of CD2831 was detected as above and quantified with a microplate fluorescence reader. ( D ) ELISA plates were coated with 10 μg/ml of purified collagens and incubated with serially diluted recombinant CD2831 proteins ranging from 15 nM to 2 µM. Binding of the proteins was detected using rabbit anti-CD2831 serum, followed by HRP-conjugated-secondary antibody. CWP25 protein was used as negative control. CD2831 CBD1 and CD2831 CBD2 represent the previously described subdomains 1 and 2.

Article Snippet: After blocking with PBS containing 0.05% Tween-20 (PBS-T) and 10% (w/v) milk (Sigma, USA), proteins were detected with rabbit anti-CD2831 serum diluted 1:1000 for 1 hour at RT, followed by the HRP-conjugated secondary antibodies (Dako, Denmark) diluted 1:2000 for 1 hour at RT.

Techniques: Binding Assay, Recombinant, Immunofluorescence, Produced, Staining, Cell Culture, Incubation, Fluorescence, Enzyme-linked Immunosorbent Assay, Purification, Negative Control