ADAM12, BioAssay(TM) ELISA Kit (Human)

Product Overview

• Article Name: ADAM12, BioAssay(TM) ELISA Kit (Human)
• Biozol Catalog Number: USB-A0859-27
• Supplier Catalog Number: A0859-27
• Alternative Catalog Number: USB-A0859-27-1
• Manufacturer: US Biological
• Category: Kits/Assays

Product Description

Human ADAM12 (a disintegrin and metalloproteinase domain-containing protein 12) belongs to the ADAM metalloproteinase family. It is a single polypeptide chain composed of multiple domains, including the signal peptide, propeptide, metalloproteinase, disintegrin, cystein-rich, and EGF-like domains, as well as the transmembrane segment and cytoplasmic tail. Human ADAM12 has two alternatively transcribed forms: a membrane-anchored long form of 110kD and a 90kD shorter secreted form lacking the transmembrane domain and the cytoplasmic tail (1-3). ADAM12 is initially synthesized as an inactive enzyme. Its latency requires the coordination of a cysteine residue in the pro domain with the Zn2+ in the active site of the catalytic domain. It is then activated by a furin-like protease in the trans-Golgi apparatus through cleavage of the pro domain before it is transported to the cell surface and secreted as the active form (4, 5). Upon activation, the pro domain remains non-covalently attached to the mature enzyme (6). Endogenous inhibitors for ADAM12 include TIMP-3 and a2-macroglobulin (7). The soluble form of ADAM12 is detectable in various body fluids, such as serum and urine (8-10). Like other metalloproteinases, ADAM12 possesses gelatinase activity. It can cleave extracellular matrix proteins, such as gelatin, type IV collagen, and fibronectin (10). Thus, ADAM12 may play important roles in cell adhesion and extracellular matrix degradation. Over-expression of ADAM12 has been observed in various types of cancer. In mouse prostate cancer, ADAM12 is highly expressed in the carcinoma-associated stroma, and it is required for tumor progression (11). In a mouse breast cancer model, ADAM12 accelerates tumor progression through increasing the apoptotic sensitivity of normal stromal cells while rendering tumor cells more resistant to apoptosis (12, 13). Additionally, it has been reported that in breast cancer patients, ADAM12 has significantly increased amounts in the urine compared to healthy controls and higher levels of urinary ADAM12 seem to correlate with more aggressive phenotypes and more advanced stages (10). ADAM12 may also participate in cell signaling. It has been shown that it can facilitate the phosphorylation of Smad2 and the stabilization of the TGF-b receptor type II (14). It is likely that ADAM12 contributes to TGF-b signaling via a mechanism independent of its protease activity. Additionally, the cytoplasmic tail of ADAM12 can directly interact with a number of cell signaling molecules, such as a-actinin, c-Src, and PI-3 kinase (15-17). Substrates for ADAM12 also include IGFBP-3 and IGFBP-5 (8). ADAM12 is abundantly expressed in the placenta. During pregnancy, its serum levels steadily increase, which correlate with the elevation of IGFBP-3 proteolysis. Thus, it has been postulated that ADAM12 may be the enzyme that is responsible for the cleavage of IGFBP-3 during pregnancy (8). Abnormal ADAM12 serum levels in pregnant women are associated with a number of neonatal and pregnancy-related disorders, such as Downs syndrome, chromosome 18 trisomy, intra-uterine fetal growth restriction, and preeclampsia (9, 18-21). The Human ADAM12 immunoassay is a 4.5 hour solid phase ELISA designed to measure human ADAM12 in cell culture supernates, serum, plasma, and urine. It contains CHO cell-expressed recombinant human ADAM12 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural ADAM12 showed linear curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring ADAM12. Sample Type: Serum/Plasma/Urine-Samples from apparently healthy volunteers were evaluated for the presence of ADAM12 in this assay. No medical histories were available for the donors used in this study. Cell Culture Supernates-U-87 MG human glioblastoma/astrocytoma cells were cultured in MEM media supp