Anti-Matrix Metalloproteinase-12, Propeptide Region Antibody Produced in Rabbit (M6057)

ANTI-MATRIX METALLOPROTEINASE-12 (MMP-12), ProductInformation PROPEPTIDE REGION Developed in Rabbit, Affinity Isolated Antibody

Product Number M 6057

Product Description more divergent metalloproteinases. All MMPs are Anti-Matrix Metalloproteinase-12 (MMP-12) is synthesized as proenzymes, and most of them are developed in rabbit using a synthetic peptide secreted from the cells as proenzymes. Thus, the corresponding to the propeptide region of human activation of these proenzymes is a critical step that MMP-12 (macrophage elastase) as immunogen. Affinity leads to extracellular matrix breakdown. isolated antigen specific antibody is obtained from rabbit anti-MMP-12 antiserum by immuno-specific MMPs are considered to play an important role in purification which removes essentially all rabbit serum wound healing, apoptosis, bone elongation, embryo proteins, including immunoglobulins, which do not development, uterine involution, angiogenesis, 4 and specifically bind to the peptide. tissue remodeling, and in diseases such as multiple sclerosis, 2, 5 Alzheimer’s, 2 malignant gliomas, 2 lupus, Rabbit Anti-MMP-12, Propeptide Region may be used arthritis, periodontis, glumerulonephritis, atherosclero- for the detection and localization of MMP-12 by sis, tissue ulceration, and in cancer cell invasion and immunoblotting. metastasis.6 Numerous studies have shown that there is a close association between expression of various Rabbit Anti-MMP-12, Propeptide Region specifically members of the MMP family by tumors and their binds to MMP-12 and does not cross-react with the proliferative and invasive behavior and metastaic other MMP family members (MMP-1, MMP-2, MMP-3, potential. MMP-9, etc). By immunoblotting, the antibody reacts with a band at 54 kDa (zymogen) but not any of the The tissue inhibitors of metalloproteinases (TIMPs) are active forms. Anti-MMP-12 recognizes the propeptide naturally occurring proteins that specifically inhibit region of latent MMP-12, which is shed during matrix metalloproteinases and regulate extracellular enzymatic activation. It does not recognize the matrix turnover and tissue remodeling by forming tight- activated forms of MMP-12; thus it can be used to binding inhibitory complexes with the MMPs. Thus, differentiate between the zymogen and the activated TIMPs maintain the balance between matrix destruction enzyme. and formation. An imbalance between MMPs and the associated TIMPs may play a significant role in the The matrix metalloproteinases (MMPs) are a family of invasive phenotype of malignant tumors. MMPs and at least eighteen secreted and membrane-bound zinc- TIMPs can be divided into two groups with respect to endopeptidases. Collectively, these enzymes can gene expression: the majority exhibit inducible degrade all the components of the extracellular matrix, expression and a small number are produced including fibrillar and non-fibrillar collagens, fibronectin, constitutively or are expressed at very low levels and laminin and basement membrane glycoproteins. In are not inducible. Among agents that induce MMP and general, a signal peptide, a propeptide, and a catalytic TIMP production are the inflammatory cytokines TNF-α domain containing the highly conserved zinc-binding and IL-1β. A marked cell type specificity is a hallmark of site characterizes the structure of the MMPs. In both MMP and TIMP gene expression (i.e., a limited addition, fibronectin-like repeats, a hinge region, and a number of cell types can be induced to make these C-terminal hemopexin-like domain allow categorization proteins). of MMPs into the collagenase, gelatinase, stomelysin 1-3 and membrane-type MMP subfamilies. MMPs contain Matrix Metalloproteinase-12 (MMP-12, macrophage the motif His-Glu-X-X-His (X represents any amino elastase) was first described in murine macrophages, acid) that binds zinc in the catalytic site, as well as later in human macrophages, and recently in other cell another zinc molecule and two calcium molecules types. MMP-12 is also known as metalloelastase and structurally. They fall within the matrixin subfamily and EC 3.4.24.65. MMP-12 degrades elastin, entactin, are EC designated 3.4.24.x. This group also contains laminin-1, fibronectin, type IV collagen as well as insulin astacin, reprolysin, and serralysin, as well as other B-chain and casein. Structurally, human MMP-12 is similar to the classical from a stimulated human cell line, an alkaline MMPs (MMP-1, MMP-3), sharing 49 % amino acid phosphatase conjugated secondary antibody, and identity. MMP-12 contains a propeptide with an BCIP/NBT as substrate. Higher antibody concentrations autoinhibitory cysteine switch site, a well-conserved may be necessary for non-human samples. zinc site, a hinge region and a hemopexin. MMP-12 lacks a transmembrane domain and a furin cleavage Note: Since cell types differ greatly in the quantity of site. The zymogen for MMP-12 is approximately 54 kDa MMP-12 produced, the conditioned media may require and is quickly activated to the 45 kDa form, which protein concentration or mitogen stimulation to visualize breaks down to a cascade of active forms, ending with the bands by immunoblotting. a 22 kDa form. In order to obtain best results and assay sensitivity in MMP-12 levels in quiescent cells and tissue are different techniques and preparations we recommend minimal (with the exception of macrophages). determining optimum working dilutions by titration Stimulated macrophages produce MMP-12. MMP-12 assay. has also been detected in osteosarcoma cells, synovial fibroblasts and lung fibroblasts. MMP-12 plays a central References role in the pathogenesis of pulmonary emphysema7 and 1. Borkakoti, N., Matrix metalloproteases: variations atherosclerotic lesions.8 In addition, MMP-12 may have on a theme. Prog. Biophy. Mol. Biol., 70, 73 (1998). beneficial functions such as modulating macrophage 2. Yong, V.W., et al., Matrix metalloproteinases and influx to inflammatory sites and limiting growth of tumor diseases of the CNS. Trends in Neuroscience, 21, metastases.9 75 (1998). The human MMP-12 gene has the chromosomal 3. Kähäri, V.M., and Saarialho-Kere, U., Matrix location of 11q22.2/22.3.10 metalloproteinases in skin. Exp. Dermatol., 6, 199 (1997). Reagent 4. Halpert, I., et al., Matrilysin is expressed by lipid- Rabbit Anti-MMP-12, Propeptide Region, is supplied in laden macrophages at sites of potential rupture in 0.01 M phosphate buffered saline, pH 7.4, containing atherosclerotic lesions and localizes to areas of 50 % glycerol and 0.1 % sodium azide. versican deposition, a proteoglycan substrate for Protein concentration is approximately 1 mg/ml. the enzyme. Proc. Natl. Acad. Sci., USA, 93, 9748 (1996). Precautions and Disclaimer 5. Chandler, S., et al., Matrix metalloproteinases, Due to the sodium azide content a material safety data tumor necrosis factor and multiple sclerosis: an sheet (MSDS) for this product has been sent to the overview. J. Neuroimmunol., 72, 155 (1997). attention of the safety officer of your institution. Consult 6. Birkedal-Hansen, H., et al., Matrix metallo- the MSDS for information regarding hazards and safe proteinases: a review. Crit. Rev. Oral. Biol. Med., 4, handling practices. 197 (1993). 7. Hautamaki, R.D., et al., Requirement for Storage/Stability macrophage elastase for cigarette smoke-induced For continuous use, store at 2 ° to 8 °C for up to six emphysema in mice. Science, 277, 2002-2004 months. For extended storage, the solution may be (1997). stored 0 °C to −20 °C. The antibody is supplied with 8. Carmeleit, O., et al., Plasmin predisposes to 50 % glycerol to prevent freezing. If slight turbidity atherosclerotic aneurysm formation by activation of occurs upon prolonged storage, clarify the solution by matrix metalloproteinases. Nature Genetics, 17, centrifugation before use. 439-444 (1997). 9. Dong, Z., et al., Macrophage-derived metallo- Product Profile elastase is responsible for the generation of A working dilution of 1:1,000 is determined by angiostatin in Lewis lung carcinoma. Cell, 88, 801- immunoblotting using a concentrated cell culture media 810 (1997). 10. Belaaouaj, A., et al., Human macrophage metallo- expression. J. Biol. Chem., 270, 14568-14575 elastase. Genomic organization, chromosomal (1995). location, gene linkage, and tissue-specific kaa 10/00

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