Fibronectin is a high molecular weight glycoprotein found on the surfaces of many cell types and present in many extracellular ﬂuids including plasma. Fibronectin binds speciﬁcally to gelatin at or around physiological pH and ionic strength.
Binding buffer: PBS: 140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.4
Elution buffer alternatives:
Fibronectin has a tendency to bind to glass. Use siliconized glass to prevent adsorption.
Wash 3 times with 2–3 column volumes of buffer, alternating between high pH (0.1 M Tris-HCl, 0.5 M NaCl, pH 8.5) and low pH (0.1 M sodium acetate, 0.5 M NaCl, pH 4.5). Re-equilbrate immediately with 3–5 column volumes of binding buffer. Remove denatured proteins or lipids by washing the column with 0.1% Triton X-100 at +37 °C for one minute. Re-equilibrate immediately with 5 column volumes of binding buffer.
Stable in all commonly used aqueous buffers.
Wash media and columns with 20% ethanol at neutral pH (use approximately 5 column volumes for packed media) and store at +4 to +8 °C.
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