Fibronectin (FN1) Rabbit Polyclonal Antibody

Specifications

Product Data
Applications	ELISA, IHC, IP, WB
Recommend Dilution	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. This product was assayed by immunoblot and found to be reactive against Fibronectin at a dilution of 1:5,000 to 1:10,000. This product was also assayed against 1.0 ?g of Fibronectin in a standard capture ELISA using Peroxidase Conjugated Streptavidin and ABTS (2,2'-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) as a substrate for 30 minutes at room temperature. A working dilution of 1:4,000 to 1:8,000 of the stock concentration is suggested for this product. For immunohistochemistry on paraffin embedded tissue dilute the product 1:50 to 1:200. Recommended Dilutions: ELISA: 1/5,000-1/20,000. Western Blot: 1/500-1/5,000. Immunoprecipitation: 1/100. Immunohistochemistry: 1/50-1/200.
Reactivity	Human
Host	Rabbit
Clonality	Polyclonal
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.
Specificity	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 most mammalian Fibronectins 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.
Formulation	0.02M Potassium Phosphate, 0.15M Sodium Chloride, pH 7.2, containing 10 mg/ml BSA (IgG and Protease free) as stabilizer and 0.01% (w/v) Sodium Azide as preservative. Label: Biotin State: Lyophilized purified Ig fraction Label: Biotinamidocaproate N-Hydroxysuccinimide Ester (BAC) Molar radio: 10-20 BAC molecules per Rabbit IgG molecule.
Reconstitution Method	Restore with 0.1 ml of deionized water or equivalent.
Concentration	lot specific
Purification	Immunoaffinity Chromatography.
Conjugation	Biotin
Storage Condition	Store vial at 2-8°C prior to restoration. Restore with deionized water (or equivalent); centrifuge product if not completely clear after standing at room temperature. This product is stable for one month at 2-8°C as an undiluted liquid. For extended storage reconstitute product with 50% glycerol instead of water and then aliquot contents and freeze at -20°C or below. Avoid repeated freezing and thawing. Dilute only prior to immediate use.
Gene Name	fibronectin 1
Database Link	Entrez Gene 2335 Human UniProt ID P02751
Background	Human fibronectin has a molecular weight of 450 kDa when purified in an intact form from plasma. Fibronectin is a glycoprotein synthesized in the liver for the circulating blood plasma form, and is synthesized by many mesenchymal cells, for the extracellular matrix form. It is composed of two similar, but not identical protein chains, which are linked by two disulfide linkages at the C-terminal end of the chains. The chains are composed of domains which have specific secondary structures linked together by regions which are especially susceptible to proteolytic action. For this reason, detection by immunoblot (western) may show considerable variability in the observed apparent molecular weights, which is predicated on the source of the fibronectin, and to what degree proteolysis has occurred. Bands approximately 225 kDa shoeld be observed after SDS-PAGE when reducing and denaturing conditions are used.
Synonyms	FN1, Cold-insoluble globulin, CIG
Reference Data

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