Fibronectin was purified from Human plasma by binding to a denatured gelatin column followed by elution with high concentrations of arginine. The eluted material was further purified by gel filtration. Immunization occurred after single-band purity was assessed by SDS-PAGE.
Anti-Fibronectin (rabbit) antibody was tested by WB, ELISA, and IHC. Assay by immunoblot was found to be reactive against Fibronectin at a dilution of 1:5,000 to 1:10,000. Assay against 1.0 µg of Fibronectin in a standard capture ELISA using Peroxidase C
Immunohistochemistry of Rabbit Anti-Fibronectin Antibody. Tissue: human kidney at pH9 at 20x and 40x. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Fibronectin antibody at 10 µg/mL for 1 h at RT. Secondary antibody: Peroxidase rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: Fibronectin is cytoplasmic. Staining: Fibronectin as precipitated brown signal (A) with purple nuclear counterstain. With corresponding negative control (B).
Immunohistochemistry of Rabbit Anti-Fibronectin Antibody. Tissue: human kidney at pH6 at 20x and 40x. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Fibronectin antibody at 10 µg/mL for 1 h at RT. Secondary antibody: Peroxidase rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: Fibronectin is cytoplasmic. Staining: Fibronectin as precipitated brown signal (A) with purple nuclear counterstain. With corresponding negative control (B).
Immunohistochemistry with rabbit anti fibronectin biotin conjugated at 20X with negative controls (right). Tissue: kidney. Fixation: FFPE buffered formalin 10% conc. Antigen retrieval: Heat, Citrate pH 6.2. Pressure Cooker (top) or EDTA pH 9.5 Pressure Cooker (bottom). Primary antibody: 2ug/ml for 1 hour room T. Secondary antibody: Streptav. Conj. HRP 10 ug/ml circa 45 min. room T. Staining: antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.
Unfolding profile of Biotin Conjugated Rabbit Anti-Fibronectin Antibody. The fluorescence signal is plotted against temperature. The vertical line indicates the Ti at 63.4C. Independently Validated byantibodies-online GmbH (p/n ABIN5596762) courtesy of NanoTemper Technologies.
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