CloneID: REGA-3G12
Antigen Long Description: The original antibody was generated by immunizing Balb/c mice with a purified gelatinase B enzyme preparation.
Origin Pub PMID: 9323036
Buffer Composition: PBS only.
Uniprot Accession No.: P14780
Specificity Statement: This antibody is specific for human gelatinase B; it recognizes the catalytic site region between Trp116 and Lys214 of the enzyme's globular aminoterminal part. A dimer scFv (with a His6 tag) variant of this antibody was found to cross-react with gelatinase A.
Application Notes (Clone): The specificity of this antibody's original format (mouse IgG1, κ) was evaluated by ELISA; it specifically bound to gelatinase B with a binding affinity of 2.1 x 10-9 M. This antibody detected both native and H2NPhAcHg-treated gelatinase B in an immunoblot, both in non-reducing and reducing conditions. Furthermore, this antibody was found to inhibit gelatinase B activity in vitro (Paemen et al., 1995; PMID: 8575432). The binding of the scFv version of this antibody to gelatinase B was confirmed in an ELISA. The original antibody and its F(ab')2 and scFv derivatives inhibited gelatinase B in vitro (Zhou et al., 1997; PMID: 9323036). The following scFv derivatives of this antibody were evaluated for their inhibitory effect toward gelatinase B: REGA-3G12 scFv with (His6) monomer (10 μM), REGA-3G12 scFv with (His6) dimer (5 μM), and REGA-3G12 scFv without a tag (1.25, 2.5, and 5 μM). They achieved 6.5%, 65.9%, 8.8%, 32.4%, and 44.4% inhibitory activity, respectively. Moreover, the scFv His-tag dimer variant was found to cross-react with gelatinase A with a 75% inhibitory activity (same antibody concentration) (Hu et al., 2004; PMID: 15104254). The original format of the antibody was used to detect and characterize the binding of this antibody to gelatinase B via immunoprecipitation (IP) and Western blot (WB), and the catalytic domain in the aminoterminal segment of gelatinase B was identified as its target (Martens et al., 2007; PMID: 17137715).
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