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Santa Cruz Biotechnology mouse monoclonal antibodies against human fibronectin

Differentiated Caco-2 cells intoxicated with TcdA and TcdB for 3, 6, or 8h in DMEM FBS-free. As a control, cells were treated with DMEM FBS-free. Unpermeabilized cells were stained for accessible <t>Fibronectin</t> or Vitronectin (acc Fn or acc Vn; green), permeabilized, and stained total Fibronectin or Vitronectin (total Fn or total Vn; red) and nuclei (blue). a, b Representative confocal microscopy images 3D projection of control cells (left) and intoxicated cells for 8h (right) immunostained for acc Fn or Vn and total Fn or Vn, below a magnified slide (XY), and the orthogonal view (XZ). Relative fluorescence intensity measured as the sum of raw intensity density/area for each z-step of accFn and total Fn, its abundance in the c apical side or d in the basal side of the cell; in the same way, the relative fluorescence intensity of Vn, its abundance in the e apical side and f the basal side of the cell. g , immunoblotting of anti- nonglucosylated Rac1 and total Rac1 of cell lysates of differentiated Caco-2 cells intoxicated with TcdA and TcdB for 3, 6, or 8 h. Nonglucosylated Rac1 was evaluated with corresponding antibodies, then the membrane was stripped, and subsequently tested for total Rac1. Western blotting is representative of 3 independent experiments. Controls were set at 100%. Error bars indicate the mean ± SEM from at least 9 fields ( n = 3). Statistical analysis was performed by Two- Way ANOVA post-Bonferroni; ns, p > 0.05; * p < 0.05. Scale bar, top panels 20µm; bottom panels 5µm.

Mouse Monoclonal Antibodies Against Human Fibronectin, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1771 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Clostridioides difficile major toxins remodel the intestinal epithelia, affecting spore adherence/internalization into intestinal tissue and their association with gut vitronectin"

Article Title: Clostridioides difficile major toxins remodel the intestinal epithelia, affecting spore adherence/internalization into intestinal tissue and their association with gut vitronectin

Journal: bioRxiv

doi: 10.1101/2025.01.29.635439

Figure Legend Snippet: Differentiated Caco-2 cells intoxicated with TcdA and TcdB for 3, 6, or 8h in DMEM FBS-free. As a control, cells were treated with DMEM FBS-free. Unpermeabilized cells were stained for accessible Fibronectin or Vitronectin (acc Fn or acc Vn; green), permeabilized, and stained total Fibronectin or Vitronectin (total Fn or total Vn; red) and nuclei (blue). a, b Representative confocal microscopy images 3D projection of control cells (left) and intoxicated cells for 8h (right) immunostained for acc Fn or Vn and total Fn or Vn, below a magnified slide (XY), and the orthogonal view (XZ). Relative fluorescence intensity measured as the sum of raw intensity density/area for each z-step of accFn and total Fn, its abundance in the c apical side or d in the basal side of the cell; in the same way, the relative fluorescence intensity of Vn, its abundance in the e apical side and f the basal side of the cell. g , immunoblotting of anti- nonglucosylated Rac1 and total Rac1 of cell lysates of differentiated Caco-2 cells intoxicated with TcdA and TcdB for 3, 6, or 8 h. Nonglucosylated Rac1 was evaluated with corresponding antibodies, then the membrane was stripped, and subsequently tested for total Rac1. Western blotting is representative of 3 independent experiments. Controls were set at 100%. Error bars indicate the mean ± SEM from at least 9 fields ( n = 3). Statistical analysis was performed by Two- Way ANOVA post-Bonferroni; ns, p > 0.05; * p < 0.05. Scale bar, top panels 20µm; bottom panels 5µm.

Techniques Used: Control, Staining, Confocal Microscopy, Fluorescence, Western Blot, Membrane

Ileal ligated loops were intoxicated for 5 h with 0.1, 0.5, 1, or 5µg of TcdB or saline as control. Then loops were removed, washed, fixed, and subjected to immunofluorescence. Unpermeabilized tissues were stained for accessible Fibronectin or Vitronectin (acc Fn or acc Vn; green), and then permeabilized and stained total Fibronectin or Vitronectin (total Fn or total Vn; red) and F-actin (grey). a - b , Representative confocal microscopy images 3D projection of control cells (left) or intoxicated loops with 5µg TcdB (right) immunostained for accessible and total Fn or Vn; right bottom, a magnified 3D projection, next to a z-stack (XY), and then magnified orthogonal view (XZ). Quantification of c, e , accessible or d, f , total Fn or Vn fluorescence intensity per cell measured in the z-projection (sum). For acc Fn or Vn, the analyzed area was Ctrl of 170,360 µm 2 ; 0.1 µg TcdB of 340,720 µm 2 ; 0.5 µg TcdB of 340,720 µm 2 ; 1 µg TcdB of 340,720 µm 2 and 5 µg TcdB of 511,080 µm 2 . n = 3 animal per group. In scatter plots, each dot corresponds to one independent cell. Dots in colors correspond to the average of each analyzed mice/field. Error bars indicate mean or mean ± SEM. Statistical analysis was performed by unpaired Student’s t test; ns, p > 0.05; * p < 0.05; ** p < 0.01; **** p < <0.0001. Scale bar 20 µm.

Figure Legend Snippet: Ileal ligated loops were intoxicated for 5 h with 0.1, 0.5, 1, or 5µg of TcdB or saline as control. Then loops were removed, washed, fixed, and subjected to immunofluorescence. Unpermeabilized tissues were stained for accessible Fibronectin or Vitronectin (acc Fn or acc Vn; green), and then permeabilized and stained total Fibronectin or Vitronectin (total Fn or total Vn; red) and F-actin (grey). a - b , Representative confocal microscopy images 3D projection of control cells (left) or intoxicated loops with 5µg TcdB (right) immunostained for accessible and total Fn or Vn; right bottom, a magnified 3D projection, next to a z-stack (XY), and then magnified orthogonal view (XZ). Quantification of c, e , accessible or d, f , total Fn or Vn fluorescence intensity per cell measured in the z-projection (sum). For acc Fn or Vn, the analyzed area was Ctrl of 170,360 µm 2 ; 0.1 µg TcdB of 340,720 µm 2 ; 0.5 µg TcdB of 340,720 µm 2 ; 1 µg TcdB of 340,720 µm 2 and 5 µg TcdB of 511,080 µm 2 . n = 3 animal per group. In scatter plots, each dot corresponds to one independent cell. Dots in colors correspond to the average of each analyzed mice/field. Error bars indicate mean or mean ± SEM. Statistical analysis was performed by unpaired Student’s t test; ns, p > 0.05; * p < 0.05; ** p < 0.01; **** p < <0.0001. Scale bar 20 µm.

Techniques Used: Saline, Control, Immunofluorescence, Staining, Confocal Microscopy, Fluorescence

Differentiated Caco-2 cells were intoxicated with 600pM of TcdA and TcdB for 8h in DMEM FBS-free media. Controls include non-intoxoicated cells in DMEM FBS-free. Supernatant was collected from untreated and intoxicated cells and subsequently utilized to resuspend C. difficile spores and incubate for 1 h at 37 °C. Spores resuspended in DMEM alone were also included as a control. Spores were washed and strained for immunofluorescence anti- fibronectin and - vitronectin. a-b, Micrographs show representative phase-contrast (phase), fibronectin and vitronectin specific immunofluorescence and fluorescence intensity profiles (Fl. int.). Representative Fl. int. were provided using 3D Surface plotter function of Fiji. c - d , Quantitative analysis of the fluorescence Fl. int. of Fn and Vn in spores Fl. Int of 600 spores. Mean ± SEM are denoted. e -g, Ileal ligated loops were intoxicated with 5µg of TcdB and 5 × 10 8 C. difficile R20291 spores for 5 h. Then loops were removed, washed, fixed, and subjected to immunofluorescence. Unpermeabilized tissues were stained for accessible Fibronectin or Vitronectin (acc Fn or acc Vn; green), and then permeabilized and stained total Fibronectin or Vitronectin (total Fn or total Vn; red) and F-actin (grey). e, g , representative 3D confocal micrograph projection reconstruction of fixed whole-mount small intestine tissue, and magnification of C. difficile spores associated with Fn or Vn. Plot profiles of fluorescence intensity of C. difficile spores (red line) and accessible Fn or Vn (green lines). f,h , quantification of spores that were positive (Fn+) or negative (Fn-) for Fn fluorescence signal in f , or positive (Vn+) or negative (Vn-) for Vn fluorescence signal in h . The average of associated and non-associated spores with f Fn or h Vn for each field. A total of ∼ 500 spores were counted per mice (n = 5 per group). GRUBB’s test was performed to identify outliers, and one point was removed in Vn. Error bars indicate mean ± S.E.M. Statistical analysis was performed by two-tailed unpaired Student’s t test; ns indicates non-significant differences. Scale bar, 20 μm.

Figure Legend Snippet: Differentiated Caco-2 cells were intoxicated with 600pM of TcdA and TcdB for 8h in DMEM FBS-free media. Controls include non-intoxoicated cells in DMEM FBS-free. Supernatant was collected from untreated and intoxicated cells and subsequently utilized to resuspend C. difficile spores and incubate for 1 h at 37 °C. Spores resuspended in DMEM alone were also included as a control. Spores were washed and strained for immunofluorescence anti- fibronectin and - vitronectin. a-b, Micrographs show representative phase-contrast (phase), fibronectin and vitronectin specific immunofluorescence and fluorescence intensity profiles (Fl. int.). Representative Fl. int. were provided using 3D Surface plotter function of Fiji. c - d , Quantitative analysis of the fluorescence Fl. int. of Fn and Vn in spores Fl. Int of 600 spores. Mean ± SEM are denoted. e -g, Ileal ligated loops were intoxicated with 5µg of TcdB and 5 × 10 8 C. difficile R20291 spores for 5 h. Then loops were removed, washed, fixed, and subjected to immunofluorescence. Unpermeabilized tissues were stained for accessible Fibronectin or Vitronectin (acc Fn or acc Vn; green), and then permeabilized and stained total Fibronectin or Vitronectin (total Fn or total Vn; red) and F-actin (grey). e, g , representative 3D confocal micrograph projection reconstruction of fixed whole-mount small intestine tissue, and magnification of C. difficile spores associated with Fn or Vn. Plot profiles of fluorescence intensity of C. difficile spores (red line) and accessible Fn or Vn (green lines). f,h , quantification of spores that were positive (Fn+) or negative (Fn-) for Fn fluorescence signal in f , or positive (Vn+) or negative (Vn-) for Vn fluorescence signal in h . The average of associated and non-associated spores with f Fn or h Vn for each field. A total of ∼ 500 spores were counted per mice (n = 5 per group). GRUBB’s test was performed to identify outliers, and one point was removed in Vn. Error bars indicate mean ± S.E.M. Statistical analysis was performed by two-tailed unpaired Student’s t test; ns indicates non-significant differences. Scale bar, 20 μm.

Techniques Used: Control, Immunofluorescence, Fluorescence, Staining, Two Tailed Test

Model of C. difficile toxin-mediated spore interactions and Bezlotoxumab effects on intestinal epithelial cells. Left panel (CDI): During C. difficile infection, toxins A and B (TcdA/TcdB) cause increased accessible (Acc) fibronectin (Fn), vitronectin (Vn), and their associated integrins (α5 and αv) on both apical and basal surfaces of intestinal epithelial cells. This leads to enhanced spore adherence and internalization, ultimately resulting in epithelial cell apoptosis. Right panel (CDI + Bezlotoxumab): Treatment with Bezlotoxumab partially neutralizes TcdB, reducing epithelial damage and spore adherence/internalization. While total Vn levels increase, total Fn levels decrease. The presence of Bezlotoxumab antibodies results in reduced spore adherence and internalization compared to untreated conditions, demonstrating partial protection against TcdB- mediated damage. Arrows indicate increased (↑) or decreased (↓) levels of proteins. A and B represent TcdA and TcdB toxins, respectively. The apical and basal sides of the epithelium are indicated.

Figure Legend Snippet: Model of C. difficile toxin-mediated spore interactions and Bezlotoxumab effects on intestinal epithelial cells. Left panel (CDI): During C. difficile infection, toxins A and B (TcdA/TcdB) cause increased accessible (Acc) fibronectin (Fn), vitronectin (Vn), and their associated integrins (α5 and αv) on both apical and basal surfaces of intestinal epithelial cells. This leads to enhanced spore adherence and internalization, ultimately resulting in epithelial cell apoptosis. Right panel (CDI + Bezlotoxumab): Treatment with Bezlotoxumab partially neutralizes TcdB, reducing epithelial damage and spore adherence/internalization. While total Vn levels increase, total Fn levels decrease. The presence of Bezlotoxumab antibodies results in reduced spore adherence and internalization compared to untreated conditions, demonstrating partial protection against TcdB- mediated damage. Arrows indicate increased (↑) or decreased (↓) levels of proteins. A and B represent TcdA and TcdB toxins, respectively. The apical and basal sides of the epithelium are indicated.

Techniques Used: Infection

Incubation:

Article Title: Stomatin-like Protein 2 Is Overexpressed in Cancer and Involved in Regulating Cell Growth and Cell Adhesion in Human Esophageal Squamous Cell Carcinoma
Article Snippet: .. Sections were incubated with the polyclonal primary antibody against SLP-2 at 1:200 dilution or mouse monoclonal antibody against fibronectin at 1:100 dilution (Santa Cruz Biotechnology) for 2 hours at 37jC or overnight at 4jC. ..

Article Title: CTGF Is Increased in Basal Deposits and Regulates Matrix Production through the ERK (p42/p44 mapk ) MAPK and the p38 MAPK Signaling Pathways
Article Snippet: Each sample containing 30 μ g of total protein of cell lysates or 20 μ L of supernatant was separated by SDS-PAGE and electroblotted to a polyvinylidene fluoride (PVDF) membrane (Bio-Rad). .. After blocking nonspecific binding with 5% skim milk, the membranes were incubated with a mouse monoclonal antibody against fibronectin (1:200; Santa Cruz Biotechnology, Santa Cruz, CA), laminin (1:200), MMP-2 (1:100), or a rabbit polyclonal antibody against p38 MAPK (1:1000), ERK1/2 (1:1000), phosphorylated forms of p38 MAPK or ERK1/2 (1:1000; Cell Signaling Technology, Danvers, MA), followed by incubation with a horseradish peroxidase–conjugated goat antibody against rabbit IgG (GE Healthcare, Buckinghamshire, UK). .. The signals were visualized with a kit (ECL kit; GE Healthcare) according to the manufacturer's protocol.

Purification:

Article Title: The Antifibrosis Effects of Peroxisome Proliferator-Activated Receptor δ on Rat Corneal Wound Healing after Excimer Laser Keratectomy
Article Snippet: .. The primary antibodies used were peptide-affinity purified goat antibody against α -SMA (1 : 100 dilution; Abgent, USA), mouse monoclonal antibody against Col3a1 (1 : 100 dilution; Santa Cruz, CA, USA), and mouse monoclonal antibody against fibronectin (1 : 100 dilution; Santa Cruz, CA, USA). .. After PBST washing, the sections were incubated with FITC-labeled rabbit anti-goat IgG (H+L) (1 : 200 dilution; Earthox, USA) or TRITC-labeled rabbit anti-mouse IgG (H+L) (1 : 200 dilution; Earthox, USA) for 90 min at room temperature.

Blocking Assay:

Binding Assay:

other:

Article Title: Berberine ameliorates renal injury in diabetic C57BL/6 mice: Involvement of suppression of SphK-S1P signaling pathway.
Article Snippet: Berberine (BBR) was previously found to have beneficial effects on renal injury in experimental diabetic rats.. However, the mechanisms underlying the effects are not fully understood.. Sphingosine kinase–Sphingosine 1-phosphate (SphK–S1P) signaling pathway has been implicated in the pathogenesis of diabetic nephropathy (DN).

Image Search Results

Journal: bioRxiv

Article Title: Clostridioides difficile major toxins remodel the intestinal epithelia, affecting spore adherence/internalization into intestinal tissue and their association with gut vitronectin

doi: 10.1101/2025.01.29.635439

Figure Lengend Snippet: Differentiated Caco-2 cells intoxicated with TcdA and TcdB for 3, 6, or 8h in DMEM FBS-free. As a control, cells were treated with DMEM FBS-free. Unpermeabilized cells were stained for accessible Fibronectin or Vitronectin (acc Fn or acc Vn; green), permeabilized, and stained total Fibronectin or Vitronectin (total Fn or total Vn; red) and nuclei (blue). a, b Representative confocal microscopy images 3D projection of control cells (left) and intoxicated cells for 8h (right) immunostained for acc Fn or Vn and total Fn or Vn, below a magnified slide (XY), and the orthogonal view (XZ). Relative fluorescence intensity measured as the sum of raw intensity density/area for each z-step of accFn and total Fn, its abundance in the c apical side or d in the basal side of the cell; in the same way, the relative fluorescence intensity of Vn, its abundance in the e apical side and f the basal side of the cell. g , immunoblotting of anti- nonglucosylated Rac1 and total Rac1 of cell lysates of differentiated Caco-2 cells intoxicated with TcdA and TcdB for 3, 6, or 8 h. Nonglucosylated Rac1 was evaluated with corresponding antibodies, then the membrane was stripped, and subsequently tested for total Rac1. Western blotting is representative of 3 independent experiments. Controls were set at 100%. Error bars indicate the mean ± SEM from at least 9 fields ( n = 3). Statistical analysis was performed by Two- Way ANOVA post-Bonferroni; ns, p > 0.05; * p < 0.05. Scale bar, top panels 20µm; bottom panels 5µm.

Article Snippet: For staining surface-accessible proteins, cells were incubated with primary antibodies (1:200 dilution) including mouse monoclonal antibodies against human fibronectin (sc8422, Santa Cruz Biotechnologies, USA), vitronectin (sc74484, Santa Cruz Biotechnologies, USA), integrin α 5 (ab78614, Abcam, USA), integrin α v (ab16821, Abcam, USA), and integrin β 1 (MAB1959Z, Millipore, USA).

Techniques: Control, Staining, Confocal Microscopy, Fluorescence, Western Blot, Membrane

Journal: bioRxiv

Article Title: Clostridioides difficile major toxins remodel the intestinal epithelia, affecting spore adherence/internalization into intestinal tissue and their association with gut vitronectin

doi: 10.1101/2025.01.29.635439

Figure Lengend Snippet: Ileal ligated loops were intoxicated for 5 h with 0.1, 0.5, 1, or 5µg of TcdB or saline as control. Then loops were removed, washed, fixed, and subjected to immunofluorescence. Unpermeabilized tissues were stained for accessible Fibronectin or Vitronectin (acc Fn or acc Vn; green), and then permeabilized and stained total Fibronectin or Vitronectin (total Fn or total Vn; red) and F-actin (grey). a - b , Representative confocal microscopy images 3D projection of control cells (left) or intoxicated loops with 5µg TcdB (right) immunostained for accessible and total Fn or Vn; right bottom, a magnified 3D projection, next to a z-stack (XY), and then magnified orthogonal view (XZ). Quantification of c, e , accessible or d, f , total Fn or Vn fluorescence intensity per cell measured in the z-projection (sum). For acc Fn or Vn, the analyzed area was Ctrl of 170,360 µm 2 ; 0.1 µg TcdB of 340,720 µm 2 ; 0.5 µg TcdB of 340,720 µm 2 ; 1 µg TcdB of 340,720 µm 2 and 5 µg TcdB of 511,080 µm 2 . n = 3 animal per group. In scatter plots, each dot corresponds to one independent cell. Dots in colors correspond to the average of each analyzed mice/field. Error bars indicate mean or mean ± SEM. Statistical analysis was performed by unpaired Student’s t test; ns, p > 0.05; * p < 0.05; ** p < 0.01; **** p < <0.0001. Scale bar 20 µm.

Techniques: Saline, Control, Immunofluorescence, Staining, Confocal Microscopy, Fluorescence

Journal: bioRxiv

Article Title: Clostridioides difficile major toxins remodel the intestinal epithelia, affecting spore adherence/internalization into intestinal tissue and their association with gut vitronectin

doi: 10.1101/2025.01.29.635439

Figure Lengend Snippet: Differentiated Caco-2 cells were intoxicated with 600pM of TcdA and TcdB for 8h in DMEM FBS-free media. Controls include non-intoxoicated cells in DMEM FBS-free. Supernatant was collected from untreated and intoxicated cells and subsequently utilized to resuspend C. difficile spores and incubate for 1 h at 37 °C. Spores resuspended in DMEM alone were also included as a control. Spores were washed and strained for immunofluorescence anti- fibronectin and - vitronectin. a-b, Micrographs show representative phase-contrast (phase), fibronectin and vitronectin specific immunofluorescence and fluorescence intensity profiles (Fl. int.). Representative Fl. int. were provided using 3D Surface plotter function of Fiji. c - d , Quantitative analysis of the fluorescence Fl. int. of Fn and Vn in spores Fl. Int of 600 spores. Mean ± SEM are denoted. e -g, Ileal ligated loops were intoxicated with 5µg of TcdB and 5 × 10 8 C. difficile R20291 spores for 5 h. Then loops were removed, washed, fixed, and subjected to immunofluorescence. Unpermeabilized tissues were stained for accessible Fibronectin or Vitronectin (acc Fn or acc Vn; green), and then permeabilized and stained total Fibronectin or Vitronectin (total Fn or total Vn; red) and F-actin (grey). e, g , representative 3D confocal micrograph projection reconstruction of fixed whole-mount small intestine tissue, and magnification of C. difficile spores associated with Fn or Vn. Plot profiles of fluorescence intensity of C. difficile spores (red line) and accessible Fn or Vn (green lines). f,h , quantification of spores that were positive (Fn+) or negative (Fn-) for Fn fluorescence signal in f , or positive (Vn+) or negative (Vn-) for Vn fluorescence signal in h . The average of associated and non-associated spores with f Fn or h Vn for each field. A total of ∼ 500 spores were counted per mice (n = 5 per group). GRUBB’s test was performed to identify outliers, and one point was removed in Vn. Error bars indicate mean ± S.E.M. Statistical analysis was performed by two-tailed unpaired Student’s t test; ns indicates non-significant differences. Scale bar, 20 μm.

Techniques: Control, Immunofluorescence, Fluorescence, Staining, Two Tailed Test

Journal: bioRxiv

Article Title: Clostridioides difficile major toxins remodel the intestinal epithelia, affecting spore adherence/internalization into intestinal tissue and their association with gut vitronectin

doi: 10.1101/2025.01.29.635439

Figure Lengend Snippet: Model of C. difficile toxin-mediated spore interactions and Bezlotoxumab effects on intestinal epithelial cells. Left panel (CDI): During C. difficile infection, toxins A and B (TcdA/TcdB) cause increased accessible (Acc) fibronectin (Fn), vitronectin (Vn), and their associated integrins (α5 and αv) on both apical and basal surfaces of intestinal epithelial cells. This leads to enhanced spore adherence and internalization, ultimately resulting in epithelial cell apoptosis. Right panel (CDI + Bezlotoxumab): Treatment with Bezlotoxumab partially neutralizes TcdB, reducing epithelial damage and spore adherence/internalization. While total Vn levels increase, total Fn levels decrease. The presence of Bezlotoxumab antibodies results in reduced spore adherence and internalization compared to untreated conditions, demonstrating partial protection against TcdB- mediated damage. Arrows indicate increased (↑) or decreased (↓) levels of proteins. A and B represent TcdA and TcdB toxins, respectively. The apical and basal sides of the epithelium are indicated.

Techniques: Infection

(A) Confluent ECs were incubated with MPO (20 nM) for the indicated times (0-120 min) and cellular levels of eNOS and MPO assessed in cell lysates by Western Blotting. (B, C) Confluent ECs on coverslips were incubated with MPO (20 nM) for 2 h, washed and then fixed with paraformaldehyde and immunostained for MPO (red) or fibronectin (green) by laser confocal fluorescence microscopy. Serial images were taken along the z-plane using the z-stack feature included in the microscopy software. Images were processed using ImageJ, where the apical, intracellular and basolateral planes were determined by referencing the DAPI-stained cell nuclei (blue).

Journal: Free radical biology & medicine

Article Title: Endothelial-Transcytosed Myeloperoxidase Activates Endothelial Nitric Oxide Synthase via a Phospholipase C-Dependent Calcium Signaling Pathway

doi: 10.1016/j.freeradbiomed.2020.12.448

Figure Lengend Snippet: (A) Confluent ECs were incubated with MPO (20 nM) for the indicated times (0-120 min) and cellular levels of eNOS and MPO assessed in cell lysates by Western Blotting. (B, C) Confluent ECs on coverslips were incubated with MPO (20 nM) for 2 h, washed and then fixed with paraformaldehyde and immunostained for MPO (red) or fibronectin (green) by laser confocal fluorescence microscopy. Serial images were taken along the z-plane using the z-stack feature included in the microscopy software. Images were processed using ImageJ, where the apical, intracellular and basolateral planes were determined by referencing the DAPI-stained cell nuclei (blue).

Article Snippet: Fixed cells were probed with a rabbit polyclonal antibody against MPO (1:300; Merck Millipore) or a mouse monoclonal antibody against fibronectin (1:100; Clone E3E, Merck Millipore), followed by the relevant labelled secondary antibody; i.e., cyanine 2-conjugated goat anti-mouse (green) or cyanine 3-conjugated donkey anti-rabbit (red) antibodies.

Techniques: Incubation, Western Blot, Fluorescence, Microscopy, Software, Staining

$(A–D) Fibronectin predominantly appeared along the laser incision site or lenticular interface. The expression was reduced over time after either ReLEx or refractive lenticule re-implantation. (E–H) Tenascin was absent along the flap interface on week 8 and 16 following ReLEx, but was present along the borders of the stromal lenticule after re-implantation. The intensity of the staining was attenuated over time. (I–L) Collagen type I was expressed uniformly in the full thickness of corneal stroma. No significant anomaly in collagen arrangement was observed in the corneas post-ReLEx and post-reimplantation. (M–P) CD18-positive cells were not seen in all post-operative corneas. Unoperated corneas were used as control. Arrowheads indicate the location of the laser incision site or lenticular interface. PR: post-ReLEx, PLR: post-lenticule re-implantation. Scale bar: 50 µm.$

Journal: PLoS ONE

Article Title: Reversible Femtosecond Laser-Assisted Myopia Correction: A Non-Human Primate Study of Lenticule Re-Implantation after Refractive Lenticule Extraction

doi: 10.1371/journal.pone.0067058

Figure Lengend Snippet: (A–D) Fibronectin predominantly appeared along the laser incision site or lenticular interface. The expression was reduced over time after either ReLEx or refractive lenticule re-implantation. (E–H) Tenascin was absent along the flap interface on week 8 and 16 following ReLEx, but was present along the borders of the stromal lenticule after re-implantation. The intensity of the staining was attenuated over time. (I–L) Collagen type I was expressed uniformly in the full thickness of corneal stroma. No significant anomaly in collagen arrangement was observed in the corneas post-ReLEx and post-reimplantation. (M–P) CD18-positive cells were not seen in all post-operative corneas. Unoperated corneas were used as control. Arrowheads indicate the location of the laser incision site or lenticular interface. PR: post-ReLEx, PLR: post-lenticule re-implantation. Scale bar: 50 µm.

Article Snippet: The following primary antibodies and the corresponding working dilution factor were used: mouse monoclonal antibody against cellular fibronectin (Millipore, Billerica, MA) diluted 1∶400; tenascin-C (Abcam, Cambridge, UK) diluted 1∶200; collagen type I (Sigma) diluted 1∶100; and CD18 (Novus Biologicals, Littleton, CO) diluted 1∶100 in the blocking solution.

Techniques: Expressing, Staining

Maternal characteristics and first trimester glycosylated fibronectin and fibronectin concentrations

Journal: Archives of Gynecology and Obstetrics

Article Title: Glycosylated fibronectin as a first trimester marker for gestational diabetes

doi: 10.1007/s00404-020-05670-8

Figure Lengend Snippet: Maternal characteristics and first trimester glycosylated fibronectin and fibronectin concentrations

Article Snippet: Normal fibronectin assay was done by coating 96-well plates with mouse monoclonal IgG antibody against human fibronectin (MAB1918, R&D Systems, Abingdon, UK).

Techniques: Sampling

Correlation of glycosylated fibronectin (SNA-based assay) and normal fibronectin (antibody-based assay). There was a high and significant correlation between the assays across control ( n = 59) and GDM ( n = 19) groups

Journal: Archives of Gynecology and Obstetrics

Article Title: Glycosylated fibronectin as a first trimester marker for gestational diabetes

doi: 10.1007/s00404-020-05670-8

Figure Lengend Snippet: Correlation of glycosylated fibronectin (SNA-based assay) and normal fibronectin (antibody-based assay). There was a high and significant correlation between the assays across control ( n = 59) and GDM ( n = 19) groups

Article Snippet: Normal fibronectin assay was done by coating 96-well plates with mouse monoclonal IgG antibody against human fibronectin (MAB1918, R&D Systems, Abingdon, UK).

Techniques: Control

a Glycosylated fibronectin concentrations in BMI subgroups of 20–25 and 30–35 kg/m 2 in the study and the control group. b Effect of smoking on concentrations of glycosylated fibronectin in the control and GDM groups. c Multiple of median (MoM) values for fibronectin and glycosylated fibronectin between control and GDM groups. d Multiple of median (MoM) values for adiponectin and glycosylated adiponectin between control and GDM groups

Journal: Archives of Gynecology and Obstetrics

Article Title: Glycosylated fibronectin as a first trimester marker for gestational diabetes

doi: 10.1007/s00404-020-05670-8

Figure Lengend Snippet: a Glycosylated fibronectin concentrations in BMI subgroups of 20–25 and 30–35 kg/m 2 in the study and the control group. b Effect of smoking on concentrations of glycosylated fibronectin in the control and GDM groups. c Multiple of median (MoM) values for fibronectin and glycosylated fibronectin between control and GDM groups. d Multiple of median (MoM) values for adiponectin and glycosylated adiponectin between control and GDM groups

Article Snippet: Normal fibronectin assay was done by coating 96-well plates with mouse monoclonal IgG antibody against human fibronectin (MAB1918, R&D Systems, Abingdon, UK).

Techniques: Control

Screening performance using the logistic regression model for fibronectin ( a ) and glycosylated fibronectin ( b ). AUC area under the curve

Journal: Archives of Gynecology and Obstetrics

Article Title: Glycosylated fibronectin as a first trimester marker for gestational diabetes

doi: 10.1007/s00404-020-05670-8

Figure Lengend Snippet: Screening performance using the logistic regression model for fibronectin ( a ) and glycosylated fibronectin ( b ). AUC area under the curve

Article Snippet: Normal fibronectin assay was done by coating 96-well plates with mouse monoclonal IgG antibody against human fibronectin (MAB1918, R&D Systems, Abingdon, UK).

Techniques:

Atropine effects on collagen and fibronectin production by HSF cells. Western immunoblot analysis of collagen type-1α1 (Col-1α1) and fibronectin in HSF lysates treated with atropine at different doses as indicated for 24 and 48 h. A representative Western immunoblot is shown at the bottom of each panel, and the quantitative analysis is reported in the histogram above. Numbers on bars indicate the percentage value with respect to the control, set at 100%. Each bar represents the average value ± SD of three different experiments: ( A ) Col-1α1 24 h; ( B ) Fibronectin 24 h; ( C ) Col-1α1 48 h; and ( D ) fibronectin 48 h. * p < 0.05 vs. CTRL. One-way ANOVA, followed by Tukey’s test.

Journal: Biomedicines

Article Title: Atropine Differentially Modulates ECM Production by Ocular Fibroblasts, and Its Ocular Surface Toxicity Is Blunted by Colostrum

doi: 10.3390/biomedicines8040078

Figure Lengend Snippet: Atropine effects on collagen and fibronectin production by HSF cells. Western immunoblot analysis of collagen type-1α1 (Col-1α1) and fibronectin in HSF lysates treated with atropine at different doses as indicated for 24 and 48 h. A representative Western immunoblot is shown at the bottom of each panel, and the quantitative analysis is reported in the histogram above. Numbers on bars indicate the percentage value with respect to the control, set at 100%. Each bar represents the average value ± SD of three different experiments: ( A ) Col-1α1 24 h; ( B ) Fibronectin 24 h; ( C ) Col-1α1 48 h; and ( D ) fibronectin 48 h. * p < 0.05 vs. CTRL. One-way ANOVA, followed by Tukey’s test.

Article Snippet: Membranes were next incubated with rabbit polyclonal antibodies against Col-1α1 (Cell Signaling, Leiden, Netherlands, Cat. No. 84336), and GAPDH (Cell Signaling, Leiden, Netherlands, Cat. No. 2118), or mouse monoclonal antibodies against fibronectin (Abcam, Cambridge, United Kingdom, Cat. No. ab6328), overnight at 4 °C followed by incubation with IgG-HRP-conjugated anti-rabbit or anti-mouse secondary antibodies (Amersham, GE Healthcare, Illinois, USA, Cat. no. NA934V) for 1 h at room temperature.

Techniques: Western Blot, Control

7MX effects on collagen and fibronectin production by HSF cells. Western immunoblot analysis of collagen type-1α1 (Col-1α1) and fibronectin in HSF lysates treated with 7MX at different doses as indicated for 24 and 48 h. A representative Western immunoblot is shown at the bottom of each panel, and the quantitative analysis is reported in the histogram above. Numbers on bars indicate the percentage value with respect to the control, set at 100%. Each bar represents the average value ± SD of three different experiments: ( A ) Col-1α1 24 h; ( B ) Fibronectin 24 h; ( C ) Col-1α1 48 h; and ( D ) fibronectin 48 h. * p < 0.05 vs. CTRL. One-way ANOVA, followed by Tukey’s test.

Journal: Biomedicines

Article Title: Atropine Differentially Modulates ECM Production by Ocular Fibroblasts, and Its Ocular Surface Toxicity Is Blunted by Colostrum

doi: 10.3390/biomedicines8040078

Figure Lengend Snippet: 7MX effects on collagen and fibronectin production by HSF cells. Western immunoblot analysis of collagen type-1α1 (Col-1α1) and fibronectin in HSF lysates treated with 7MX at different doses as indicated for 24 and 48 h. A representative Western immunoblot is shown at the bottom of each panel, and the quantitative analysis is reported in the histogram above. Numbers on bars indicate the percentage value with respect to the control, set at 100%. Each bar represents the average value ± SD of three different experiments: ( A ) Col-1α1 24 h; ( B ) Fibronectin 24 h; ( C ) Col-1α1 48 h; and ( D ) fibronectin 48 h. * p < 0.05 vs. CTRL. One-way ANOVA, followed by Tukey’s test.

Techniques: Western Blot, Control

Atropine and 7MX effects on collagen and fibronectin production by HCOF cells. Western immunoblot analysis of collagen type-1α1 (Col-1α1) and fibronectin in HCOF lysates treated with atropine or 7MX at 100 μM for 48 h. A representative Western immunoblot is shown at the bottom of each panel, and the quantitative analysis is reported in the histogram above. Numbers on bars indicate the percentage value with respect to the control, set at 100%. Each bar represents the average value ± SD of three different experiments: ( A ) Col-1α1 48 h; and ( B ) fibronectin 48 h. * p < 0.05 vs. CTRL. One-way ANOVA, followed by Tukey’s test.

Journal: Biomedicines

Article Title: Atropine Differentially Modulates ECM Production by Ocular Fibroblasts, and Its Ocular Surface Toxicity Is Blunted by Colostrum

doi: 10.3390/biomedicines8040078

Figure Lengend Snippet: Atropine and 7MX effects on collagen and fibronectin production by HCOF cells. Western immunoblot analysis of collagen type-1α1 (Col-1α1) and fibronectin in HCOF lysates treated with atropine or 7MX at 100 μM for 48 h. A representative Western immunoblot is shown at the bottom of each panel, and the quantitative analysis is reported in the histogram above. Numbers on bars indicate the percentage value with respect to the control, set at 100%. Each bar represents the average value ± SD of three different experiments: ( A ) Col-1α1 48 h; and ( B ) fibronectin 48 h. * p < 0.05 vs. CTRL. One-way ANOVA, followed by Tukey’s test.

Techniques: Western Blot, Control

Review

mouse monoclonal antibodies against human fibronectin (Santa Cruz Biotechnology)

Structured Review

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1) Product Images from "Clostridioides difficile major toxins remodel the intestinal epithelia, affecting spore adherence/internalization into intestinal tissue and their association with gut vitronectin"

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