Invitrogen™ Aggrecan Monoclonal Antibody (BC-3)

Mouse Monoclonal Antibody

Purified recombinant Aggrecan Interglobular Domain (IGD) protein (Amino acids T331-G458 with C-term His-tag, expressed in E. coli), was used to validate this antibody. The recombinant protein contains cleavage sites for aggrecanases (E373-A374 in aggrecan) and matrix metalloproteinases (N341-F342 in aggrecan). The recombinant protein was cleaved in vitro using recombinant Aggrecanase. Degradation of IGD and accumulation of cleaved fragments was visualized via MEMCode reversible protein staining, followed by Western Blot using the BC-3 antibody for the specific detection of the aggrecan neoepitope. The antibody will also recognize full length IGD, as shown by ELISA, however with significantly lower affinity. MA3-16888 has successfully been used in ELISA, immunohistochemistry (frozen and FFPE) and Western blot applications. This antibody may be used to immunostain formalin-fixed, paraffin-embedded sections, as well as alcohol-fixed frozen sections and un-fixed snap frozen sections. Samples must be deglycosylated using 0.01 Units Chondroitinase ABC, 0.01 Units Keratanase and 0.0001 Units Keratanase II per 10 μg S-GAG of non-deglycosylated aggrecan for optimal epitope recognition. MA3-16888 detects aggrecan in human, bovine, feline, canine, guinea pig, equine, porcine, rabbit, rat, and sheep samples.