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. Does not react with mouse samples.
Aggrecan also known as cartilage-specific proteoglycan core protein (CSPCP) or chondroitin sulfate proteoglycan 1 is a protein that is encoded by the ACAN gene. The encoded protein is an integral part of the extracellular matrix in cartilaginous tissue and it withstands compression in cartilage. The human form of the protein is 2316 amino acids long (~ 250 kDa) and can be expressed in multiple isoforms due to alternative splicing. Aggrecan exhibits a bottlebrush structure, in which chondroitin sulfate and keratan sulfate chains are attached to an extended protein core. The synthesis and degradation of aggrecan are being investigated for their roles in cartilage deterioration during joint injury, disease, and aging. It contains three interglobular domains (IGDs), G1, G2, and G3 that are involved in aggregation, hyaluronan binding, cell adhesion, and chondrocyte apoptosis. Aggrecan fragments from articular cartilage are released into the synovial fluid at all stages of human osteoarthritis as a result of cleavage by aggrecanase and MMP activity. Antibodies BC-3 and BC-4 can be used to detect these neo epitopes of Aggrecanase (ARGSVXXX) and MMP (XXXVDIPEN), respectively.
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Protein Aliases: AGC1; AGCAN; aggrecan; aggrecan 1; aggrecan 1 (chondroitin sulfate proteoglycan 1, large aggregating proteoglycan, antigen identified by monoclonal antibody A0122); Aggrecan core protein; aggrecan epidermal growth factor-like domain-1; aggrecan interglobular domain; aggrecan, structural proteoglycan of cartilage; cartilage aggregating proteoglycan; Cartilage-specific proteoglycan core protein; Chondroitin sulfate proteoglycan 1; Chondroitin sulfate proteoglycan core protein 1; CSPCP; CSPG1; CSPGCP; large aggregating proteoglycan; large aggregating proteoglycan, antigen; MSK16; SEDK
Gene Aliases: ACAN; Agc; AGC1; AGCAN; BOS_20116; CSPCP; CSPG1; CSPGCP; MSK16; SEDK