منابع مشابه
Subcellular western blotting of single cells
Although immunoassays are the de facto standard for determining subcellular protein localization in individual cells, antibody probe cross-reactivity and fixation artifacts remain confounding factors. To enhance selectivity while providing single-cell resolution, we introduce a subcellular western blotting technique capable of separately assaying proteins in the 14 pL cytoplasm and 2 pL nucleus...
متن کاملWestern Blotting
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...
متن کاملMicrofluidic Western blotting.
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...
متن کاملImmunoprecipitation and western blotting
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...
متن کاملImmunofluorescent protein detection in Western blotting
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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ژورنال
عنوان ژورنال: Nature Protocols
سال: 2016
ISSN: 1754-2189,1750-2799
DOI: 10.1038/nprot.2016.089