Preparative elution of proteins from nitrocellulose membranes after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Parekh, Bharat; Mehta, Hina; West, Melanie; Montelaro, R. C.

doi:10.1016/0003-2697(85)90631-1

Cited by 78 publications

(19 citation statements)

References 19 publications

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“…protein was eluted from the filter by incubating in 40 percent acetonitrile, 0. 1M anmmomum acetate (pH 8.9) for 3 hours at 370C (26). The protein was lyophilized and subjected to partial amino acid hydrolysis, and phosphoamino acids were separated by two-dimensional thin-layer electrophoresis (27).…”

mentioning

confidence: 99%

A Yeast Gene That Is Essential for Release from Glucose Repression Encodes a Protein Kinase

Celenza

Carlson

1986

Science

680

438

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The SNF1 gene plays a central role in carbon catabolite repression in the yeast Saccharomyces cerevisiae, namely that SNF1 function is required for expression of glucose-repressible genes. The nucleotide sequence of the cloned SNF1 gene was determined, and the predicted amino acid sequence shows that SNF1 encodes a 72,040-dalton polypeptide that has significant homology to the conserved catalytic domain of mammalian protein kinases. Specific antisera were prepared and used to identify the SNF1 protein. The protein was shown to transfer phosphate from adenosine triphosphate to serine and threonine residues in an in vitro autophosphorylation reaction. These findings indicate that SNF1 encodes a protein kinase and suggest that protein phosphorylation plays a critical role in regulation by carbon catabolite repression in eukaryotic cells.

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mentioning

confidence: 99%

A Yeast Gene That Is Essential for Release from Glucose Repression Encodes a Protein Kinase

Celenza

Carlson

1986

Science

680

438

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“…The proteins were transferred after electrophoresis to nitrocellulose membranes, excised, and eluted from the membranes by overnight incubation at 37°C in 50% pyridine-100 mM ammonium acetate (pH 8.9) as described previously (30). When phosphoamino acid analysis was performed on total proteins, these proteins were isolated as described previously (39).…”

mentioning

confidence: 99%

Differential protein phosphorylation in induction of thyroid cell proliferation by thyrotropin, epidermal growth factor, or phorbol ester.

Contor¹,

Lamy²,

Lecocq³

et al. 1988

Mol. Cell. Biol.

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Protein phosphorylation was studied in primary cultures of thyroid epithelial cells after the addition of different mitogens: thyrotropin (TSH) acting through cyclic AMP, epidermal growth factor (EGF), or 12-O-tetradecanoylphorbol-13-acetate (TPA). EGF or TPA increased the phosphorylation of five common polypeptides. Among these, two 42-kilodalton proteins contained phosphotyrosine and phosphoserine with or without phosphothreonine. Their characteristics suggested that they are similar to the two 42-kilodalton target proteins for tyrosine protein phosphorylation demonstrated in fibroblasts in response to mitogens. No common phosphorylated proteins were detected in TSH-treated cells and in EGF- or TPA-treated cells. The differences in the protein phosphorylation patterns in response to TSH, EGF, and TPA suggested that the newly emerging cyclic AMP-mediated mitogenic pathway is distinct from the better known growth factor- and tumor promoter-induced pathways.

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“…The desired band was located by staining the gel with 4 M sodium acetate. Thereafter, a horizontal strip containing the antigen band was excised and treated with a solution (30 ml) containing 6 M urea, 50 mM phosphate buffer, pH 7, and 20 mM EDTA at 37°C for 18 h. Afterwards, the extract obtained was filtered, dialyzed against water, and finally liophylized (12).…”

Section: Purification Of the 38-kda Antigenmentioning

confidence: 99%

Rapid screening test for tuberculosis using a 38-kDa antigen fromMycobacterium tuberculosis

Rosales-Borjas

Zambrano-Villa

Elinos

et al. 1998

J. Clin. Lab. Anal.

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Preparative elution of proteins from nitrocellulose membranes after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Cited by 78 publications

References 19 publications

A Yeast Gene That Is Essential for Release from Glucose Repression Encodes a Protein Kinase

A Yeast Gene That Is Essential for Release from Glucose Repression Encodes a Protein Kinase

Differential protein phosphorylation in induction of thyroid cell proliferation by thyrotropin, epidermal growth factor, or phorbol ester.

Rapid screening test for tuberculosis using a 38-kDa antigen fromMycobacterium tuberculosis

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