Open Access Open Access Volume 4 • Issue 8 • 1000203 J Chromat Separation Techniq ISSN: 2157-7064 JCGST, an open access journal Identification of Proteins in Tissue Fluids of the sea mussel Isognomon alatus Angel Alberto Justiz Vaillant 1 *, Suzette Curtello 2 , Monica Smikle 3 , Brian Wisdom 4 , Wayne Mohammed 1 , Sehlule Vuma 1 , Geeta Kurhade 5 , Norma McFarlane-Anderson 2 , Chalapathi Rao 1 , Shivnarine Kissoon 1 1 Pathology and Microbiology Unit, Department of Para-Clinical Sciences,The University of the West Indies, St. Augustine, Trinidad and Tobago 2 Department of Basic Medical Sciences, The University of the West Indies, Mona, Jamaica 3 Department of Microbiology, The University Hospital of West Indies, Mona, Jamaica 4 School of Biology and Biochemistry, Medical Biology Centre, The Queen’s University of Belfast, Belfast, Ireland 5 Physiology Section, Department of Pre-clinical Sciences, The University of the West Indies, St Augustine, Trinidad and Tobago *Corresponding author: Angel Justiz Vaillant, Department of Para-Clinical Sciences, The University of the West Indies, St. Augustine, Trinidad and Tobago, Tel: +868-736-0440; Fax: +868-663-3797; E-mail: avail4883@gmail.com Received August 29, 2013; Accepted September 29, 2013; Published October 04, 2013 Citation: Justiz Vaillant AA, Farlane-Anderson M, Smikle M, Wisdom B, Mohammed W, et al. (2013) Identifcation of Proteins in Tissue Fluids of the sea mussel Isognomon alatus. J Chromatograph Separat Techniq 4: 203. doi:10.4172/2157- 7064.1000203 Copyright: © 2013 Justiz Vaillant AA, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract The aim of this preliminary study was to investigate the presence of proteins in the tissue fuids of the sea mussel Isognomon alatus. Protein extraction was done by the chloroform-cold ethanol technique. Immunization for production of antibody to be used as reagents in Western blotting, assessment of the protein concentration by the Bradford method, protein characterization by native polyacrylamide gel electrophoresis (PAGE) were also performed as a part of the methodology of this study. The results showed a protein content of 65 mg/ml in tissue fuids and a protein band approximately of 220 kDa in PAGE that was further confrmed by the Western blotting. Future work should investigate the structure and function of the proteins separated from the tissue fuids and we considered it as a limitation of this investigation. The sea bivalve literature is scanty. However the limitation of this work we still can conclude that there are high molecular weight proteins in large concentrations in tissue fuids of the sea mussel Isognomomalatus. Keywords: Sea mussel; Isognomon alatus; Polyacrylamide gel electrophoresis (PAGE); Protein Introduction The mussel Isognomon alatus a bivalve mollusc. This species inhabits warm marine littoral habitats [1]. Despite the economic importance of this mussel, which is used for human consumption in many countries, there are no reports on the protein content of the tissue fuids of this organism. High protein content has been described in other species of mussels, e.g. the horse mussel Modiolus modiolus [2]. One approach to understanding the nutritional value of the Isognomon alatus was to determine the protein presence and concentrations in extracellular fuids of the mussel. In addition we studied the presence of Salmonella spp. In the mussel, which is eaten row by the Jamaican and other countries populations, constitute a risk to human health. Other microorganisms have been isolated from the mussel I. alatus. Tis research paper resumes the work done in a moderate amount of I. alatus (40 mussels), which is known for the Jamaican population as a “fat oyster” and considered a sexual stimulant. I. alatus protein concentration in extracellular fuids and Salmonella contamination were assessed in this study. Te scientifc literature of I. alatus is limited to a few studies and that was a hindrance to our study that has to present a little information about the mussel but with this work we are contributing to expand the knowledge of the sea mussel I. alatus and stimulating to other scientists to research on this topic. Material and Methods Collection of tissue fuids from the Isognomon alatus Te extracellular fuid secreted from the I. alatus was collected in a 50 ml sterile plastic tube and it was placed in sterile plastic bags and transported to the laboratory in ice packs, for analysis. Protein extractions from the tissue fuids of the mussel Isognomonalatus One volume of chloroform (15 ml) was added to 15 ml of tissue fuids from 10 mussels and the mixture centrifuged at 1500×g (low gravity) for 5 min at 4°C. Te clear supernatant (top layer) was separated from lipids (bottom layer) and treated with cold ethanol (100%) and centrifuged for another 5 min at 1500×g and 4°C. A large white pellet (protein extract) (5 ml) was resuspended in one volume (5 ml) of phosphate bufered saline (PBS), and dialyzed against BPS, pH=7.4 for 24 hours at 4°C, with 3 bufer changes and stored at -20°C until further analysis. Te protein concentration was assessed by Bradford method Duplicate aliquots 0.5 mg/ml of bovine serum albumin (BSA) (5, 10, 15, 20 µl) was added to micro centrifuge tubes and the volume in each tube brought to 100 µl with 0.15 M NaCl. Two blank tubes were also prepared. One ml of Coomassie brilliant blue solution was added to each tube and each vortexed and lef to stand for 2 min at room temperature (RT). Te absorbance readings were taken at 595 nm against a 0.15 M NaCl blank using a 1 cm path-length microcuvette. A standard curve was then constructed by plotting a graph of absorbance against protein concentration and the protein concentration was determined from the standard curve. Justiz Vaillant et al., J Chromatograph Separat Techniq 2013, 4:9 DOI: 10.4172/2157-7064.1000203 Research Article Journal of Chromatography Separation Techniques J o u r n a l o f C h r o m a t o g r a p h y & S e p a r a t i o n T e c h n i q u es ISSN: 2157-7064