A rapid method for fibronectin purification on nitrocellulose membranes suitable for tissue culture.
We have developed a simple method for plasma fibronectin purification based on the well-known gelatin binding property of fibronectin. In this procedure we immobilize the melted gelatin to nitrocellulose membranes; these are then used to affinity-purify the fibronectin from the plasma sample. The fibronectin is eluted from the membrane by treatment with 8 M urea. The procedure described here gives a yield of up to 60% (from presumed fibronectin concentration) and the fibronectin obtained is homogeneous in SDS-PAGE and biologically active, as assessed by a cell migration assay. The method is rapid, simple, inexpensive, does not require the use of chromatographic equipment and is suitable for tissue culture applications.
Chifflet
Silvia
S
Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Gral. Flores 2125, 11800 Montevideo, Uruguay. schiffle@mednet.org.uy
Bolatto
Carmen
C
Tolosa
Susana
S
D003101:Collodion / Q000737:chemistry*; D005353:Fibronectins / Q000097:blood / Q000302:isolation & purification* / Q000378:metabolism; D005780:Gelatin; D005898:Glass; D006801:Humans; D013997:Time Factors; D046509:Tissue Culture Techniques / Q000295:instrumentation*; D014508:Urea
D005353:Fibronectins; D014508:Urea; D005780:Gelatin; D003101:Collodion