ELISA, Immunohistochemistry, Immunoprecipitation, Western Blot. Anti-Fibronectin antibodies have been used for indirect trapping ELISA for quantitation of antigen in serum using a standard curve, for immunoprecipitation and for western blotting for highly sensitive qualitative analysis. Rockland's anti-Fibronectin detects intact fibronectin (Invitrogen, Cat. No. 33016-015) by western blot after digestion by Matrix Metalloproteinase-3 (MMP-3) overnight at 37° C. Separation was performed using a 4-12% Tris-Glycine gel. Under these conditions a sizeable, dark band at ~220 kDa representing the undigested fibronectin, as well as many, smaller bands representing the variably sized fragments resulting from fibronectin digestion by MMP-3 were noted. For immunohisto-chemistry paraffin embedded tissue preparation is recommended.
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Description
This product has been prepared by immunoaffinity chromatography using immobilized antigens followed by extensive cross-adsorption against human serum proteins and collagen and non-collagen extracellular matrix proteins to remove any unwanted specificities. Typically less than 1% cross reactivity against other extracellular matrix proteins was detected by ELISA against purified standards. This antibody reacts with human Fibronectin and has negligible cross-reactivity with Type I, II, III, IV, V or VI Collagens or Laminin. Non-specific cross reaction of anti-Fibronectin antibodies with other human serum proteins or non-Fibronectin extracellular matrix proteins is negligible.
Immunogen
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.