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Antigen retrieval in cryostat tissue sections and cultured cells by treatment with sodium dodecyl sulfate (SDS) | Histochemistry and Cell Biology
Description:
A simple method for antigen retrieval in tissue sections and cell cultures is described. Because many antibodies recognize denatured proteins on western blots, but are poorly reactive by immunocytochemistry, the effect of applying sodium dodecyl sulfate (SDS) to cryostat sections of tissues and to cell cultures prior to immunostaining was examined. In many cases, a 5-min pretreatment with 1% SDS produced a dramatic increase in staining intensity by indirect immunofluorescence. Among the antibodies tested that showed a positive effect of SDS were an anti-Na/K-ATPase monoclonal antibody, an anti-AE1/2 anion exchanger polyclonal antipeptide antibody, a monoclonal anti-caveolin antibody, and an anti-rab4 monoclonal antibody. In other cases, including antibodies against gp330, aquaporin 1, and aquaporin 2, no effect of SDS was detected. The results show that SDS treatment can be used as a simple method of antigen retrieval in cryostat sections and on cultured cells. In some cases, antigens were not detectable without pretreatment with SDS.
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google, scholar, pubmed, article, brown, cell, antigen, kidney, usa, retrieval, cells, sds, sections, antibodies, antibody, tissue, rat, renal, sci, membrane, water, privacy, cookies, content, sulfate, alper, monoclonal, access, protein, expression, proc, natl, acad, biol, publish, search, cryostat, treatment, sodium, dodecyl, immunocytochemistry, effect, gluck, physiol, microwave, channel, analysis, data, information, log,
Topics {βοΈ}
anti-na/k-atpase monoclonal antibody formalin-fixed paraffin-embedded tissues anti-rab4 monoclonal antibody monoclonal anti-caveolin antibody sodium dodecyl sulfate month download article/chapter western blot methodologies constitutively water-permeable segments periodate-lysine paraformaldehyde fixative alan stuart-tilley vasopressin-regulated water channel formalin-fixed tissue sections ae2 anion exchanger stuart-tilley ak alper sl monoclonal antibody privacy choices/manage cookies full article pdf renal brush border cell biology aims massachusetts general hospital related subjects renal proximal tubules chip28 water channel aqp2 water channel colchicine-induced redistribution distinctive glycoprotein composition chip28 water channels beth israel hospital regulated membrane expression european economic area original paper published cell cultures prior detergent-based procedure distinctive coated pit low-temperature embedding harvard medical school renal intercalated cells conditions privacy policy selectively amplified expression immunohistochemical staining based cryostat tissue sections microwave oven heating microwave antigen retrieval accepting optional cookies western blots check access rothberg kg endocytosis receptor gp330 caveolae membrane coats
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- Severs NJ (1988) Caveoli: static inpocketings of the plasma membrane, dynamic vesicles, or just plain artifact?
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headline:Antigen retrieval in cryostat tissue sections and cultured cells by treatment with sodium dodecyl sulfate (SDS)
description:A simple method for antigen retrieval in tissue sections and cell cultures is described. Because many antibodies recognize denatured proteins on western blots, but are poorly reactive by immunocytochemistry, the effect of applying sodium dodecyl sulfate (SDS) to cryostat sections of tissues and to cell cultures prior to immunostaining was examined. In many cases, a 5-min pretreatment with 1% SDS produced a dramatic increase in staining intensity by indirect immunofluorescence. Among the antibodies tested that showed a positive effect of SDS were an anti-Na/K-ATPase monoclonal antibody, an anti-AE1/2 anion exchanger polyclonal antipeptide antibody, a monoclonal anti-caveolin antibody, and an anti-rab4 monoclonal antibody. In other cases, including antibodies against gp330, aquaporin 1, and aquaporin 2, no effect of SDS was detected. The results show that SDS treatment can be used as a simple method of antigen retrieval in cryostat sections and on cultured cells. In some cases, antigens were not detectable without pretreatment with SDS.
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Western Blot
Monoclonal Antibody
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headline:Antigen retrieval in cryostat tissue sections and cultured cells by treatment with sodium dodecyl sulfate (SDS)
description:A simple method for antigen retrieval in tissue sections and cell cultures is described. Because many antibodies recognize denatured proteins on western blots, but are poorly reactive by immunocytochemistry, the effect of applying sodium dodecyl sulfate (SDS) to cryostat sections of tissues and to cell cultures prior to immunostaining was examined. In many cases, a 5-min pretreatment with 1% SDS produced a dramatic increase in staining intensity by indirect immunofluorescence. Among the antibodies tested that showed a positive effect of SDS were an anti-Na/K-ATPase monoclonal antibody, an anti-AE1/2 anion exchanger polyclonal antipeptide antibody, a monoclonal anti-caveolin antibody, and an anti-rab4 monoclonal antibody. In other cases, including antibodies against gp330, aquaporin 1, and aquaporin 2, no effect of SDS was detected. The results show that SDS treatment can be used as a simple method of antigen retrieval in cryostat sections and on cultured cells. In some cases, antigens were not detectable without pretreatment with SDS.
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