50% glycerol, 0.1 M NaCl, 0.01 M sodium phosphate at pH 7.5.
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Immunogen affinity purified
Specific antibodies were absorpted by incubation with Sepharose-bound Guinea Pig immunoglobulins.
Specific antibodies were eluted by acidic buffer at pH 2.5 followed by neutralisation and dialysis.
After repeated binding with immobilized Guinea Pig immunoglobulins a minimum of 65% protein bound.
The purified polyclonal was conjugated to fluorescein isothiocyanate isomer I followed by gel-filtration and ion-exchange chromatography to clear unbound conjugate.
F/P ratio is 3-5 mol Fluorescein per 1 mol of rabbit Ig. There is low background fluorescence of normal tissue in contrast to specific staining of sections treated with Guinea Pig anti-human fibronectin antibodies