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Thanks for your reply about ab14421. From your reply it is not clear to me the source of your peptide. I have tested direct ELISA using Recombinant human beta Defensin-1 (36 aa) from Cell Sciences, Canton, MA 02021, Catalog No. CRB501B. I have coated the wells of a microtiter plate with the above Recombinant human beta Defensin-1 peptide 50ng/well (100uL of 500ng/mL) overnight at 4 oC. After washing and blocking with 5% BSA-PBS I added Rabbit polyclonal to beta defensin-1 (your product ab14421, Lot No. 70564; dilution range: 1:100 to 1:100000). Again after washing and binding with Goat anti-rabbit IgG-HRP conjugate (Santa Cruz Biotech, sc-2004, dilution 1:10000) and color development with TMB substrate solution (KPL) I did not see much of color development above background. I want to try again ELISA exactly using your protocol. I will appreciate if you will inform me the source of human beta defensin-1 peptide you used in your ELISA and the range of dilution of Rabbit polyclonal to beta defensin-1 antiserum. Thanks!
Asked on Jan 25 2005
Thank you for your email. We source ab14421 from outside Abcam and the originator used a synthetic peptide with the sequence: Human b-Defensin 1 (aa 28-34) (YRGKAKCACMG) (underlined amino acids differ from the original sequence, they were added or substituted for technical reasons). They used the antibody at a dilution of 1:1000. If you need further assistance, please let me know.
Answered on Jan 26 2005