Carbonic anhydrases (CAs) are enzymes that catalyse the reversible hydration of CO2 to bicarbonate. MAbs (15A4 and 4A6) demonstrated a strong and specific immunostaining of CA XII in human tissue specimens. BMS-536924 Flow cytometry analysis of 5 human tumor cell lines BMS-536924 with the MAb 15A4 revealed its immunoreactivity with cellular CA XII. In conclusion, the MAbs raised against recombinant catalytic domain of CA XII recognize cellular CA XII and represent a promising diagnostic tool for the immunodetection of CA XII-expressing cells. 1. Introduction The H+ + HCO3 ?). So far, 15 different human CAs are identified that BMS-536924 differ within their enzymatic properties, subcellular localization, and cells distribution. Enzymatically energetic human being CAs are either cytosolic (CA I, CA II, CA III, CA VII, and CA XIII), membrane bound (CA IV, CA IX, CA XII, and CA XIV), mitochondrial (CA VA, and CA VB), or secretory (CA VI). CAs are indicated in a variety of cells and cells such as for example erythrocytes, gastrointestinal system, reproductive system, the nervous program, kidney, lung, pores and skin, eye, and muscle tissue [1C6]. The main function of CAs may be the transportation of CO2 in a variety of metabolizing cells. CAs are also involved in other physiological functions: pH regulation, ion transport, bone resorption and secretion of gastric juice, cerebrospinal fluid, and pancreatic juice [4]. Recent studies indicate that at least two of CA transmembrane isozymesCA IX and XIIare associated with human cancers. CA IX is a well-recognized tumor marker as it is overexpressed in many types of tumors and is found in only a few normal tissues [2]. There are increasing evidences on the potential of CA XII as a new tumor marker. CA XII is a transmembrane protein with an extracellular catalytic domain. CA XII overexpression has been demonstrated in human renal cell carcinoma [7] as well as in brain, colorectal, breast, gastrointestinal, ovarian, and pancreatic cancers [6]. CA XII is Mouse monoclonal to GFAP also expressed in the normal human kidney, colon, lung, brain, prostate, ovary, and testis [1, 8]. The expression of CA IX and CA XII is induced under hypoxic conditions through hypoxia inducible factor-1. Hypoxia and consequent acidosis of tumor microenvironment are principal features of many types of solid cancers. Both CA IX and CA XII promote tumor growth and survival through pH maintenance [6, 8C10]. Recent studies suggest that the functions of CA IX and CA XII related to tumor growth and metastasis, as well as their membrane-associated localization, make these enzymes promising targets for specific therapies. Sulfonamides represent the main group of the specific CA chemical inhibitors [11, 12]. Two monoclonal antibodies (MAbs) against CA IX were generated as specific immunological tools for clinical detection and therapy. Their diagnostic value has been confirmed by immunohistochemistry and radiolabeled monoclonal antibody imaging [9]. Recently, the importance of CA XII as a serodiagnostic marker for lung cancer has been demonstrated [13]. The MAb 6A10 raised against CA XII expressed in lung cancer cells has been shown to inhibit CA XII enzymatic activity and tumor growthin vitro[14]. The aim of the current study was to develop new monoclonal antibodies (MAbs) against human recombinant CA XII and evaluate their diagnostic potential. We have demonstrated that the MAbs raised against recombinant catalytic domain of CA XII BMS-536924 recognize cellular CA XII and are valuable reagents for its immunodetection in human tumor tissue specimens. 2. Materials and Methods 2.1. Production of Recombinant Carbonic Anhydrases Recombinant extracellular area of individual CA XII spanning amino acidity (aa) residues from 30 to 291 was portrayed inE. colicells and purified as referred to.

Carbonic anhydrases (CAs) are enzymes that catalyse the reversible hydration of

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