beta 2 Defensin antibody (ab9871)

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review by Barbara Fernie-King submitted on 22 October 2003
Application:	ELISA
beta 2 Defensin antibody (ab9871)
Additional Data
Notes	We have successfully used your goat anti-hBD-2 (ab9871) in a sandwich ELISA. Virtually no non-specific binding of the antibody was observed. We did find it necessary to use an alternative blocking protein (hydrolysed casein) instead of gelatine which we normally use, in order to prevent the target recombinant hBD-2 (Peprotech) from sticking to the uncoated/blocked control wells. The plate was coated with a protein secreted from Group A Streptococci at 1.5 ug/ml, then blocked with hydrolysed casein. Recombinant hBD-2 was added at 3 ug/ml and bound peptide detected using ab9871 diluted 1/1000 followed by alk. phos. monoclonal anti-goat Ig and pNPP. We were very pleased with this result as an ELISA performed in the other orientation had proved negative, although we knew from functional assays that hBD-2 bound to our Streptococcal protein. These results will be added to a paper which we are about to submit for publication.
Dilution notes	Goat anti-hBD2 at 1/1000 Alk. phos. monoclonal anti-goat Ig (from Sigma) 1/1200