Western Blotting: Sample Preparation to Detection
نویسندگان
چکیده
منابع مشابه
Western blotting: sample preparation to detection.
Western blotting is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide (denaturing conditions) or by the 3-D structure of the protein (native/ non-denaturing conditions). The proteins are then transferred to a membrane (typically nitrocel...
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This report describes the detailed procedures for Western blot analysis using fluorescent antibodies. After electrophoresis and subsequent electroblotting, the fluorescent-labeled antibodies were visible upon ultraviolet illumination of the polyvinylidene fluoride (PVDF) membranes and could then be photographed to give an accurate record of the blots. Fluorescent labeling allows for photographi...
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Histological Analysis and Immunohistochemistry Renal sections (3 μm thick) were stained with periodic acid-Schiff (PAS) and examined under light microscopy. Glomerulosclerosis index was semiquantitatively graded as 0 to 4+; vascular injury score as 0 to 3+; and tubulointerstitial injury score as 0 to 5+, according to the criteria reported previously. Ultrastructure of glomerular podocytes was a...
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Rapid, quantitative Western blotting is a long-sought bioanalytical goal in the life sciences. To this end, we describe a Western blotting assay conducted in a single glass microchannel under purely electronic control. The μWestern blot is comprised of multiple steps: sample enrichment, protein sizing, protein immobilization (blotting), and in situ antibody probing. To validate the microfluidic...
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SKG or BALB/c T cells (3.0 £ 10) were stimulated for 72 h with plate-bound anti-CD3 mAb (2C11) in the presence or absence of plate-bound anti-CD28 mAb in RPMI-1640 medium supplemented with 10% fetal calf serum and 50 mM 2-mercaptoethanol. Cells were also stimulated with TPA (1.4 ng ml) and ionomycin (0.14 mM). KJ1.26þ T cells from DO or DO.SKG mice were stimulated with ovalbumin (323–339) pepti...
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ژورنال
عنوان ژورنال: Journal of Visualized Experiments
سال: 2010
ISSN: 1940-087X
DOI: 10.3791/2359