Anti-Fibronectin Rabbit Monoclonal Antibody

Western blot analysis of Fibronectin using anti-Fibronectin antibody (M00564-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human U-87MG whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: rat liver tissue lysates,
Lane 4: mouse liver tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Fibronectin antigen affinity purified monoclonal antibody (Catalog # M00564-3) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:500 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Fibronectin at approximately 272 kDa. The expected band size for Fibronectin is at 272 kDa.

IHC analysis of Fibronectin using anti-Fibronectin antibody (M00564-3).
Fibronectin was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-Fibronectin Antibody (M00564-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Fibronectin using anti-Fibronectin antibody (M00564-3).
Fibronectin was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-Fibronectin Antibody (M00564-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

rFBA and fibronectin interaction analysis by Far-WB and Surface plasmon resonance (SPR) analysis. A Far-WB analysis of rFBA with fibronectin. The first lane: PVDF membrane with transferred rFBA protein incubated with anti- rFBA antibody as a positive control; the second lane: PVDF membrane with transferred rFBA protein incubated with fibronectin and anti-fibronectin antibody; the third lane: PVDF membrane with transferred BSA (negative control) incubated with fibronectin and anti-fibronectin antibody. Protein bands were visualized using ECL substrate. B Sensorgrams depict the binding of immobilised fibronectin to rFBA. Increasing concentrations of rFBA (5, 10, 25, 50 and 100 μg/mL) were injected at a flow rate of 30 μL/min for 180 s over immobilised fibronectin. The arrow indicates the end of the injection period, at which point dissociation of rFBA from fibronectin can be observed. RU resonance units.
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Representative immunofluorescence images of fibronectin. Fluorescent micrographs of tenocytes after 2 and 5 days of culture on the surface of a culture plate and acellular amniotic membrane. Tenocyte nucleus shape observed under a fluorescence microscope (A,D,G,J) . Fibronectin presented positive after the fluorescent FITC mark was observed under a fluorescence microscope (B,E,H,K) . Tenocyte nucleus and fibronectin merging (C,F,I,L) . The corresponding semi quantitative analysis of fibronectin fluorescence intensity in panels (M,N) (scale bar = 50 um, n = 5, * P < 0.05).
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Fluorescence images of tenocytes after 5 days of culture on the surface of a culture plate (control group) and acellular amniotic membrane (amnion group). Tenocytes presented a clear cytoskeleton, good biocompatibility with acellular amniotic membrane, and even distribution on the surface of the materials, showing better growth activity than the control group (A,B) . Cell viability was measured by CCK-8, and the proliferation curve of tenocytes in the control and amniotic membrane groups was drawn (C) . Western blot assay for collagen I, fibronectin, TGF-β1, and bFGF expression in the tenocytes of the control and amnion groups for 1 week (D,E) . * p < 0.05; ** p < 0.01; and *** p < 0.001.
Index in PubMed under a CC BY license. PMID: 32478059