Direct comparison of two commercially available computer programs for analysing DNA fingerprinting gels. Seward RJ, Ehrenstein B, Grundmann HJ, Towner KJ (1997). Two-dimensional gel electrophoretic analysis of vectorially labeled surface proteins of human spermatozoa. Naaby-Hansen S, Flicklinger C, Herr JC (1997). Image-Pro ®®Plus Version 1.0 for Windows ™, Reference Manual, Media Cybernetics Inc. Reproducibility of digital image analysis for measuring corneal haze after myopic photorefractive keratectomy. Maldondo M, Arnau V, Martinez-Costa R, Navea A, Mico F, Cisneros A, Menezo J (1997). A muscle hypertrophy condition in lamb (callipyge): Characterization of effects on muscle growth and meat quality traits. Koohmaraie M, Shackelford SD, Wheeler TL, Lonergan SM, Doumit ME (1995). A simplified method of analysis of cell-conditioned medium for Insulin-like Growth Factor-I (IGF-I) activity. Krabbenhoft EA, O'Reilly BA, Shultz K, Chen Y, Stewart NT, Dodson MV (1997). In: Protein Functionality in Food Systems, New York: Marcel Dekker Inc, pp 79-119. Protein separation and analysis of certain skeletal muscle proteins: Principles and Techniques. Huff-Lonergan EJ, Beekman DD, Parrish Jr FC (1994). Evaluation of satellite cell cultures by computer/video imaging enhancement: An undergraduate research project. Howard JH, Vierck J, Howell S, Dodson MV (1993). Wound status evaluation using color image processing. ![]() Hansen G, Sparrow E, Kokate J, Leland K, Iaizzo P (1997). Protein Blotting: A guide to transfer and detection. Collectively, the results of these two studies suggest that specific proteins may be evaluated by using relatively inexpensive image analysis software systems via pixel quantification of electronic images.īio-Rad Laboratories (1996). A second image analysis program, Alpha Imager™ 2000, was then used to define the boundaries of protein bands, assess pixel number and density, and to obtain final numerical data for quantifying α-Actinin on the WB. In the second procedure, WB were scanned with a ScanJet 3c® flat bed scanner and their backgrounds were clarified using Image-Pro® Plus. Dot blots corresponding to a linear concentration range from 10 to 300 ng IGF-Iwere assessed by this method. After the DB were developed and dried, the images were digitized using an HP Deskscan II® flat bed scanner, exported into Image-Pro® Plus and analyzed by taking the combined mean of 45° and 135° sample lines drawn through each dot. In the first procedure, known IGF-I samples were dotted on nitrocellulose membranes using a vacuum manifold. All rights reserved.Inexpensive computer imaging technology was used to assess levels of insulin-like growth factor-I (IGF-I) on dot blots (DB) and α-Actinin on Western blots (WB). The proposed method represents an affordable, accurate and reproducible approximation that could be used in the presence of limited resources availability.īackground subtraction Densitometry ImageJ Quantification Western blot.Ĭopyright © 2018 Elsevier B.V. Use two different software tor quantification. Optimize quantification using well established antibodies and sample lysates. In this work, we describe the use of office scanner coupled with the ImageJ software together with a new image background subtraction method for accurate Western blot quantification. 7,19 Quantification Densitometry analysis to target protein using afferent software : Inaccurate quantification to protein to interest, poor image quality, and signal saturation. Furthermore, the use of semi-quantitative software as ImageJ (Java-based image-processing and analysis software) is clearly increasing in different scientific fields. Due to the technical knowledge required for densitometry analysis together with the resources availability, standard office scanners are often used for the imaging acquisition of developed Western blot films. Western blot quantification method constitutes a critical step in order to obtain accurate and reproducible results. ![]() It constitutes a multistep method that allows the detection and/or quantification of proteins from simple to complex protein mixtures. Since its first description, Western blot has been widely used in molecular labs.
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