Nucleosomal array or H1‐nucleosomal array samples were incubated with the desired concentration of 30X MgCl2 for 30 min at room temperature and fixed with 0.1% glutaraldehyde overnight on ice. The DNA concentration was 0.215 mg/ml. Samples (10 μl drops) were deposited on freshly glow‐discharged formvar‐ and carbon‐coated copper grids for 2 min, either with no dilution or at 1:20 and 1:40 dilutions. Excess sample was removed from the grids by blotting. The grids were successively stained for 2 min with 2% uranyl acetate, sample buffer, and 1.5% phosphotungstic acid, with blotting in between each. The grids were then examined and photographed using either a JEOL JEM‐2000 EX II transmission electron microscope operated at 100 kV and captured on film, or a JEOL JEM‐1400 transmission electron microscope equipped with an Orius model 832.J76VV0 (Gatan, Inc.) digital camera and operated at 100 kV. Images were collected at microscope magnifications from 30,000 to 300,000. Negatives were scanned at 1,200 dpi using an Epson Perfection V700 photo scanner and Adobe Photoshop. Images of the grids were processed using ImageJ for figures. Magnified images of the oligomer interiors were obtained by cropping and rescaling the initial images in order to make fine details more apparent.
Perfection v700
Manufactured by Adobe
The Perfection V700 is a high-performance photo scanner designed for professional use. It features a maximum optical resolution of 6400 dpi and can handle a variety of film and document formats, including 35mm slides and negatives, medium format film, and reflective documents up to 8.5 x 11.7 inches. The scanner incorporates advanced technologies to deliver accurate color reproduction and image detail.
Automatically generated - may contain errors
Lab products found in correlation
Jem 2000 ex 2 transmission, by JEOL (1 mentions)
Anti mouse igg hrp, by Cell Signaling Technology (1 mentions)
Polyvinylidene fluoride (pvdf), by Merck Group (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Anti rabbit igg hrp, by Cell Signaling Technology (1 mentions)
Anti traf6, by Cell Signaling Technology (1 mentions)
Anti tak1, by Abcam (1 mentions)
Anti trif, by Abcam (1 mentions)
3 protocols using perfection v700
1
Negative Staining of Nucleosomal Arrays
2
Western Blot Analysis of Signaling Proteins
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Whole-cell protein extracts were resolved by SDS-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride (PVDF; Millipore, Billerica, MA). Blots were probed with anti-NFX1-123 (1:1000), anti-p53 (1:1000, Calbiochem, San Diego, CA), (anti-TRAF6 (1:500; Cell Signaling, Danvers, MA), anti-TRIF (1:1000, Abcam, Cambridge, MA), anti-TAK1 (1:500, Abcam, Cambridge, MA), anti-TAB1 (1:500, Abcam, Cambridge, MA), anti-TAB2 (1:500, Abcam, Cambridge, MA), and anti-GAPDH (1:100,000; Abcam, Cambridge, MA) primary antibodies. The secondary antibodies used were anti-mouse IgG HRP (1:10,000; Cell Signaling, Danvers, MA) or anti-rabbit IgG HRP (1:5,000, Cell Signaling, Danvers, MA). The rabbit polyclonal anti-NFX1-123 antibody was generously provided by Dr. Ann Roman. Animals were immunized with peptide composed of amino acids 1102–1120 of NFX1-123. All films were scanned using Epson Perfection V700 and imported using Adobe Photoshop.
3
Proteome Analysis of Sesbania Nodule Bacteroids
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Two-dimensional gel electrophoresis of Sesbania nodule bacteroid proteins were performed as described earlier [76 (link)]. Coomassie stained gels were destained with multiple changes of distilled water to remove background. The gels were imaged with an Epson Perfection V700 scanner controlled through Adobe Photoshop. Images were analyzed for proteome differences using Delta2D (v. 4.4.1) image analysis software.
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