FN1 Antibody

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.

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.

Store vial at 4° C prior to opening. This product is stable at 4° C as an undiluted liquid. Dilute only prior to immediate use. For extended storage mix with an equal volume of glycerol, aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing.

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. Biorbyt's anti-Fibronectin detects intact fibronectin 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 immunohistochemistry paraffin embedded tissue preparation is recommended.