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Imagemaster 2d platinum software version 7

Manufactured by GE Healthcare
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Imagemaster 2D Platinum Software Version 7.0 is a medical imaging software developed by GE Healthcare. It is designed to provide image processing and analysis capabilities for healthcare professionals.

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Lab products found in correlation

Ipgphor 2, by GE Healthcare (1 mentions) Ettan daltsix electrophoresis system, by GE Healthcare (1 mentions) Imagequant las 500, by GE Healthcare (1 mentions)

4 protocols using imagemaster 2d platinum software version 7

1

2D Gel Electrophoresis of Proteins

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Total proteins (500 μg) were loaded onto GE Healthcare 24 cm IPG gel strips (pH 4–7). The isoelectric focusing (IEF) of the acidic range IPG strips (pH 4–7) was carried out according to the manufacturer’s instructions of IPGPhor II (GE Healthcare, USA) at 20 °C for a total of 65 kVh40 (link). Two-dimensional SDS-PAGE gels (12.5% linear gradient) were ran on an Ettan Daltsix electrophoresis system (GE Healthcare, USA). The procedure was set as 2.5 W per gel for 30 min, followed by 12 W per gel for 4–5 h. After electrophoresis, the protein gels were stained for spot detection using silver nitrate, as previously described41 (link).
Gel images were analyzed using the Imagemaster 2D Platinum Software Version 7.0 (GE Healthcare). Spot detection was carried out using the software with the values of the parameters smooth, minimum area, and saliency set to 2, 15, and 8, respectively. Manual spot editing, such as spot deletion, splitting, and merging was performed. The determined relative spot intensities were subsequently used for statistical analysis38 (link).
Luan H., Shen H., Pan Y., Guo B., Lv C, & Xu R. (2018). Elucidating the hypoxic stress response in barley (Hordeum vulgare L.) during waterlogging: A proteomics approach. Scientific Reports, 8, 9655.
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2

Protein Expression Analysis by 2DE Gels

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2DE gels (silver stained) were scanned using Imaging Densitometer GS690 (Bio-Rad Laboratories, Hercules, CA, USA). Expression level of proteins was calculated in terms of percentage of volume contribution using the Image Master 2D Platinum software, version 7.0 (GE Healthcare Biosciences, Uppsala, Sweden) by selecting the particular spot of the 2DE, matching it with the same spot of another replicate (of the same time point) for which a matchset has been already created using the software. Cut-off parameters for this analysis were: Smooth—2; Saliency—1; Min area—5 (Jayapalan et al., 2012 (link); Jayapalan et al., 2013 (link)).
Subramanian P., Jayapalan J.J., Abdul-Rahman P.S., Arumugam M, & Hashim O.H. (2016). Temporal regulation of proteome profile in the fruit fly, Drosophila melanogaster. PeerJ, 4, e2080.
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3

Proteome Profiling of Wheat Flag Leaves

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Total proteins of wheat flag leaves from 10, 15, 20, and 25 DPA in three biological replicates were extracted according to [72 (link)]. Two-dimensional electrophoresis (2-DE) was used to separate the differentially accumulated protein (DAP) spots based on [73 (link)]. After 2-DE electrophoresis, all gels were stained with Coomassie Brilliant Blue (R-250:G-250 = 4:1). The ImageMaster 2D Platinum Software Version 7.0 (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) was used to analyze the gel images, and the spots with significant and biological reproducible changes (protein abundance variation at least 2-fold, Student’s t-test, p < 0.05) were considered as DAP spots.
Zhu D., Zhu G., Zhang Z., Wang Z., Yan X, & Yan Y. (2020). Effects of Independent and Combined Water-Deficit and High-Nitrogen Treatments on Flag Leaf Proteomes during Wheat Grain Development. International Journal of Molecular Sciences, 21(6), 2098.
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4

2-DE Gel Image Analysis Protocol

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The ImageQuant™ LAS500 (GE Healthcare Bioscience, Uppsala, Sweden) was used to visualize and store the silver-stained 2-DE gel images. Detection, matching, and quantification of the distinct protein spots were performed using the Image Master 2D Platinum Software, version 7.0 (GE Healthcare Bioscience, Uppsala, Sweden). Normalization was subsequently performed to evaluate proteins that were differentially expressed in the serum by correcting the spot quantification values and gel-to-gel variation unrelated to expression changes. This was achieved by taking into account total densities from the gel images (i.e., raw quantity of each gel spot was divided by the total quantity of all spots within the gel). The percentage of a given protein taken against total spot volume of all proteins (including the unresolved peptides) in the gel was calculated to give a percent of volume contribution (vol%) (Chen et al., 2008 (link)). All protein concentration values are presented as means of percentage volume (% volume) ± standard deviations (SD). Statistical analysis was done using the Student’s t-test to analyse differences between controls and patients. A p-value of less than 0.05 (p < 0.05) was considered to be statistically significant.
Kerishnan J.P., Mohammad S., Alias M.S., Mu A.K., Vaithilingam R.D., Baharuddin N.A., Safii S.H., Abdul Rahman Z.A., Chen Y.N, & Chen Y. (2016). Identification of biomarkers for periodontal disease using the immunoproteomics approach. PeerJ, 4, e2327.
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