Journal: Veterinary Research

Article Title: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) moonlights as an adhesin in Mycoplasma hyorhinis adhesion to epithelial cells as well as a plasminogen receptor mediating extracellular matrix degradation

doi: 10.1186/s13567-021-00952-8

Figure Lengend Snippet: Binding of the rGAPDH protein to different ECM components in ELISA experiment . Microtiters plate was coated with Matrigel, fibronectin, collagen or laminin solution. Various concentrations of rGAPDH or BSA were added and detected by anti-His-tag monoclonal antibody. For detecting the binding to vitronectin, microtiters plate was coated with rGAPDH or BSA. Various concentrations of vitronectin were added and detected by anti-vitronectin monoclonal antibody. * P < 0.05, ** P < 0.01, compared with the negative control (BSA).

Article Snippet: The bound vitronectin was detected by rabbit anti-vitronectin monoclonal antibody (1:2000; Boster, China), followed by goat anti-rabbit IgG (1:10 000).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Negative Control

Journal: Journal of biomolecular screening

Article Title: Screening and Identification of a Phage Display Derived Peptide That Specifically Binds to the CD44 Protein Region Encoded by Variable Exons.

doi: 10.1177/1087057115608604

Figure Lengend Snippet: Figure 1. Schematic diagram of the structure of the CD44 gene, the subtractive panning method, and the results. (A) CD44s was encoded by the standard exons (S1–S10). Between S5 and S6, variant exons (V3–V10) are spliced to form the variant protein isoforms, CD44v3-v10. (B) The phage peptide library was incubated with bovine serum albumin and CD44s prior to being screened against CD44v3-v10. Thus, only phages that bound to the CD44 variant region would be selected. (C) With each cycle of panning, the recovery ratio (calculated as the ratio of output phages/input phages) increased.

Article Snippet: Competitive Inhibition ELISA CD44v3-v10 (10 μg/mL) was immobilized on an ELISA plate and incubated with mouse anti-human CD44s antibody (AbD Serotec) or goat anti-CD44v3-v10 polyclonal antibody (AbD Serotec) at various concentrations for 1 h at 37 °C.

Techniques: Variant Assay, Incubation

Journal: Journal of biomolecular screening

Article Title: Screening and Identification of a Phage Display Derived Peptide That Specifically Binds to the CD44 Protein Region Encoded by Variable Exons.

doi: 10.1177/1087057115608604

Figure Lengend Snippet: Figure 2. Evaluation of the binding affinity and selectivity of phage clones. (A) Binding of phage clones to CD44v3-v10 (black column), CD44s (striped column), and bovine serum albumin (white column) presented as the optical density value at 450 nm. URps (unrelated phages randomly selected from the original library) and Tris-buffered saline served as the negative and blank controls, respectively. Data are presented as the mean ± SD; four measurements were made for each data point. (B) Western blotting for CD44v3-v10 and CD44s in SGC-7901 and HEK- 293 cells. GAPDH was used as a loading control. (C) CV-1 phage and unrelated phage (negative control) binding to gastric cancer SGC- 7901 cells and human embryonic kidney HEK-293 cells. Cell nuclei were stained with 4,6-diamidino-2- phenylindole. Cells were visualized using a fluorescence microscope. Scale bar, 50 µm.

Article Snippet: Competitive Inhibition ELISA CD44v3-v10 (10 μg/mL) was immobilized on an ELISA plate and incubated with mouse anti-human CD44s antibody (AbD Serotec) or goat anti-CD44v3-v10 polyclonal antibody (AbD Serotec) at various concentrations for 1 h at 37 °C.

Techniques: Binding Assay, Clone Assay, Saline, Western Blot, Control, Negative Control, Staining, Fluorescence, Microscopy

Journal: Journal of biomolecular screening

Article Title: Screening and Identification of a Phage Display Derived Peptide That Specifically Binds to the CD44 Protein Region Encoded by Variable Exons.

doi: 10.1177/1087057115608604

Figure Lengend Snippet: Figure 3. Binding of the CV-1 peptide to the CD44v3-v10 protein. (A) Peptide competitive inhibition of corresponding phage binding to CD44v3-v10. For the CV-1 phage, the inhibition ratio increased as the concentration of the peptide competitor rose. (B) Binding of CV-1 to CD44v3-v10 (solid line) and CD44s (dashed line) was presented as the optical density value at 450 nm at different concentrations. A scrambled peptide (dotted line) served as the control. (C) Binding curve of CV-1 to CD44v3-v10 and Scatchard analysis of the data. Calculated Kd = 0.46 ± 0.05 µM. Triplicate detections were performed independently. The data are presented as the mean ± SD.

Article Snippet: Competitive Inhibition ELISA CD44v3-v10 (10 μg/mL) was immobilized on an ELISA plate and incubated with mouse anti-human CD44s antibody (AbD Serotec) or goat anti-CD44v3-v10 polyclonal antibody (AbD Serotec) at various concentrations for 1 h at 37 °C.

Techniques: Binding Assay, Inhibition, Concentration Assay, Control

Journal: Journal of biomolecular screening

Article Title: Screening and Identification of a Phage Display Derived Peptide That Specifically Binds to the CD44 Protein Region Encoded by Variable Exons.

doi: 10.1177/1087057115608604

Figure Lengend Snippet: Figure 4. Competitive inhibition of CV-1 binding to CD44v3-v10 by antibodies. (A) For CV-1 (solid line), the inhibition ratio increased gradually with the concentration of preincubated anti–CD44v3-v10 antibody. A scrambled peptide (dotted line) served as a negative control. (B) The binding of both CV-1 (solid line) and the control peptide (dotted line) to CD44v3-v10 was not inhibited by an anti-CD44s antibody. Triplicate detections were performed independently, and the data are presented as the mean ± SD.

Article Snippet: Competitive Inhibition ELISA CD44v3-v10 (10 μg/mL) was immobilized on an ELISA plate and incubated with mouse anti-human CD44s antibody (AbD Serotec) or goat anti-CD44v3-v10 polyclonal antibody (AbD Serotec) at various concentrations for 1 h at 37 °C.

Techniques: Inhibition, Binding Assay, Concentration Assay, Negative Control, Control