The Proteome Profiler Mouse Cytokine Array Panel A (R&D Systems, Minneapolis, MN, USA) was used to profile cytokines in serum samples according to the manufacturer’s protocol. The array membranes were imaged by Amersham Imager 680 RGB (Cytiva Life Science). Pixel densitometry analysis in each spot of the array was carried out using ImageJ Software version 1.8.0_172 (https://imagej.nih.gov/ij/ ).
Amersham imager 680 rgb
Manufactured by Cytiva
Sourced in Germany
The Amersham Imager 680 RGB is a versatile imaging system designed for a range of life science applications. It utilizes RGB LED technology to capture high-quality images of protein gels, Western blots, and other samples. The system provides a simple and efficient way to visualize and analyze various biological samples.
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5 protocols using amersham imager 680 rgb
1
Profiling Cytokines in Mouse Serum
2
Profiling Cytokines in Mouse Serum
3
Tissue Protein Extraction and Immunoblotting
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Tissue lysates of the lung and liver from an untreated female monkey were prepared with T‐PER Tissue Protein Extraction Reagent (Thermo Fisher Scientific). The equivalent amounts of protein were separated by SDS‐PAGE and transferred to membranes. The membranes were incubated with the cathepsin B Ab (0.25 μg/mL) or a mouse monoclonal anti‐proliferating cell nuclear antigen Ab (0.3 μg/mL, clone PC10; Agilent Technologies) for 2 hours at room temperature, then with biotin‐labelled secondary Abs for 1 hour at room temperature. The signals were detected with Amersham Imager 680 RGB (Cytiva).
4
Protein Concentration Determination
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The total protein concentration in the samples was determined using the Bradford colorimetric microplate assay (Bio-Rad, Hercules, CA, USA) and absorbances were obtained at a wavelength of 595 nm using a spectrophotometer (Amersham Imager 680 RGB, Cytiva Europe GmbH, Breisgau, Germany).
5
Glycoprotein Visualization Protocol
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Glycoproteins were transferred to a polyvinylidene fluoride (PVDF) membrane and then blocked with 10 mM Tris-buffered saline pH 8 containing 0.05% Tween-20 and 3% BSA (TBS-T-BSA) at 4 °C overnight with constant rotation. Membranes were incubated for 1 h and 30 min with 1 of the 4 biotinylated lectins diluted at a concentration of 1 μg/mL in TBS-T-BSA supplemented with 1 mM CaCl2, 1 mM MnCl2,1 mM MgCl2, 1 mM ZnCl2 (TBA-T-BSA ions), followed by 5 washes in TBS-T. The membrane was then incubated with horseradish peroxidase-conjugated streptavidin (Vector Laboratories) diluted 1:5000 in TBS-T-BSA for 1 h in the dark, with rotation. After 5 washes in TBS-T, glycoproteins were visualized using an ECL immunoblot detection system (Amersham Imager 680 RGB, Cytiva Europe GmbH, Breisgau, Germany).
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