Use

ADAM-10, also known as MADM or Kuzbanian (Kuz), was first described as a membrane-associated protease that degraded myelin basic protein in the brain. A Drosophila mutant named Kuzbanian (Kuz), with defects in Notch signaling of neuronal development, was also shown to be dependent on ADAM-10. Other groups studying Alzheimer's disease identified ADAM-10 as an alpha secretase that cleaves amyloid plaque protein (APP), and other investigators showed that ADAM-10 could act as a "sheddase", releasing TNF-a. These disparate groups later identified ADAM-10 as a member of the metalloproteinase family with disintegrin domain (ADAMs), and showed that ADAM-10 also contains an EGF-like domain, transmembrane domain, and a Cytoplasmic domain. Later works describe ADAM-10 in the lung, heart, brain, kidney, and a wide range of tissues. A member of the metalloproteinase family containing disintegrin-like domains (ADAMs), ADAM-10 contains the canonical HExxHxxxxxH zinc metalloproteinase motif, and has been shown to be proteolytically active. ADAM-10 from bovine kidney was shown to have type-IV collagenolytic activity, making ADAM-10 a "gelatinase." ADAM-10 is efficiently inhibited by the endogenous MMP inhibitors TIMP-1 and TIMP-3, but not by TIMP-2 and TIMP-4. The full length ADAM10 sequence codes for an 748 amino acid protein, with a predicted mass is 84.142 kD. Glycosylation and the cyteine-rich regions make the protein run at 98 kD on reduced SDS PAGE, and 60-58 kD when Furin processed. A smaller 691 amino acid sequence for ADAM-10, lacking the transmembrane domain, has been reported, with a predicted molecular weight of 77.633 kD. ADAM-10 is thought to be membrane-anchored under normal conditions, so cell or tissue lysates are a good source for Western blots. RP4ADAM-10 recognizes the amino end of active ADAM-10, and can detect the latent and active forms of the enzyme. A recommended starting concentration for Western blots is 1:1,000 when using colorimetric substrates such as BCIP/NBT, and 1:5,000 for chemiluminescent substrates. Higher concentrations of antibody may be needed for samples from more distantly related species. EDTA/EGTA treatment of tissues or lysates is required to see latent zymogen.

FOR RESEARCH USE ONLY. NOT FOR USE IN HUMANS.

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