Thermo Fisher Scientific Aggrecan Monoclonal Antibody (BC-3)

Applications and Tested Dilutions

Product Specifications

Product Specific Information

Purified recombinant Aggrecan Interglobular Domain (IGD) protein (amino acids T331–G458 with C-terminal 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 were visualized via MEMCode reversible protein staining, followed by Western blot using the BC-3 antibody for specific detection of the aggrecan neoepitope.

The antibody also recognizes full-length IGD (shown by ELISA) but with significantly lower affinity.
MA3-16888 has been successfully used in ELISA, immunohistochemistry (frozen and FFPE), and Western blot applications.
This antibody may be used to immunostain formalin-fixed, paraffin-embedded sections, alcohol-fixed frozen sections, and un-fixed snap-frozen sections.

For optimal epitope recognition, samples must be deglycosylated using:

• 0.01 Units Chondroitinase ABC
• 0.01 Units Keratanase
• 0.0001 Units Keratanase II per 10 µg S-GAG of non-deglycosylated aggrecan

MA3-16888 detects aggrecan in human, bovine, feline, canine, guinea pig, equine, porcine, rabbit, rat, and sheep samples.

Target Information

Aggrecan, also known as cartilage-specific proteoglycan core protein (CSPCP) or chondroitin sulfate proteoglycan 1, is encoded by the ACAN gene.
It is a major component of the extracellular matrix in cartilage, providing resistance to compression.
The human protein (~250 kDa, 2316 amino acids) exists in multiple isoforms due to alternative splicing.

Aggrecan exhibits a bottlebrush structure with chondroitin sulfate and keratan sulfate chains attached to an extended core protein.
Its synthesis and degradation are key processes in cartilage deterioration during joint injury, disease, and aging.

Aggrecan contains three interglobular domains (G1, G2, G3) involved in aggregation, hyaluronan binding, cell adhesion, and chondrocyte apoptosis.
Fragments of aggrecan released into synovial fluid during osteoarthritis result from cleavage by aggrecanase and MMP activity.

Antibodies BC-3 and BC-4 detect neoepitopes of Aggrecanase (ARGSVXXX) and MMP (XXXVDIPEN), respectively.

For Research Use Only.
Not for use in diagnostic procedures.
Not for resale without express authorization.