The MBP-domain IV fusion protein is used to raise SPARC-specific antibodies. The protein is purified by amylose affinity chromatography and dialyzed into 20 mM Tris-HCl(pH 8), 50 mM NaCl. MBP-domain IV (0.2 mg in Freund's complete adjuvant) is injected subcutaneously into a rabbit. The rabbit is boosted 2 weeks later with the same amount of protein in Freund's incomplete adjuvant. The first immune bleed is collected 2 weeks after the boost and has significant anti-MBP-domain IV reactivity by immunoblotting.
ost-1