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I want to buy a mouse monoclonal antibody to aggrecan ARGxxx Cat no. ab3773. I wang to use it for immunohistochemistry for aggrecan of rattus chondrocyte. I want to konw whether I must treat the chondrocytes with digestion mixture ( Chondroitnase ABC Sigma, Keratanase Seikagaku and keratanase II Seikagaku ) as recommended before immunohistochemistry or western-blot.
Asked on May 24 2007
Thank you for your enquiry. The following reference gives details of the protocol used with this antibody in immunohistochemistry: Roberts S et al. Matrix metalloproteinases and aggrecanase: their role in disorders of the human intervertebral disc. Spine 25:3005-13 (2000). PubMed: 11145811 I have added the relevant protocol to our online datasheet for your convenience and include below the following publications which have all used this product. This product has been used in: Karsdal MA et al. Induction of increased cAMP levels in articular chondrocytes blocks matrix metalloproteinase-mediated cartilage degradation, but not aggrecanase-mediated cartilage degradation. Arthritis Rheum 56:1549-58 (2007). PubMed: 17469134 Little CB et al. Matrix metalloproteinases are involved in C-terminal and interglobular domain processing of cartilage aggrecan in late stage cartilage degradation. Matrix Biol 21:271-88 (2002). PubMed: 12009333 Flannery CR et al. Autocatalytic cleavage of ADAMTS-4 (Aggrecanase-1) reveals multiple glycosaminoglycan-binding sites. J Biol Chem 277:42775-80 (2002). PubMed: 12202483 Malfait AM et al. Inhibition of ADAM-TS4 and ADAM-TS5 prevents aggrecan degradation in osteoarthritic cartilage. J Biol Chem 277:22201-8 (2002). PubMed: 11956193 Roberts S et al. Matrix metalloproteinases and aggrecanase: their role in disorders of the human intervertebral disc. Spine 25:3005-13 (2000). PubMed: 11145811 Caterson B et al. Mechanisms involved in cartilage proteoglycan catabolism. Matrix Biol 19:333-44 (2000). PubMed: 10963994 Billington CJ et al. An aggrecan-degrading activity associated with chondrocyte membranes. Biochem J 336 ( Pt 1):207-12 (1998). PubMed: 9806902 Buttle DJ et al. "Aggrecanase" activity is implicated in tumour necrosis factor alpha mediated cartilage aggrecan breakdown but is not detected by an in vitro assay. Mol Pathol 50:153-9 (1997). PubMed: 9292151 Hughes CE et al. Monoclonal antibodies that specifically recognize neoepitope sequences generated by 'aggrecanase' and matrix metalloproteinase cleavage of aggrecan: application to catabolism in situ and in vitro. Biochem J 305 ( Pt 3):799-804 (1995). PubMed: 7531436 This clone has been used in: Molecular Approaches to Osteoarthritis Research. Abstracts of the Kennedy Institute of Rheumatology Symposium, London, United Kingdom, April 18-20, 2004. Scand J Rheumatol Suppl :1-41 (2005). PubMed: 15864865 Roberts S et al. Matrix turnover in human cartilage repair tissue in autologous chondrocyte implantation. Arthritis Rheum 44:2586-98 (2001). PubMed: 11710715 Aplin JD et al. Human endometrial MUC1 carries keratan sulfate: characteristic glycoforms in the luminal epithelium at receptivity. Glycobiology 8:269-76 (1998). PubMed: 9451036
回复于 May 24 2007