Cloning of cell-specific secreted and surface proteins by subtractive antibody screening

To identify and clone genes that encode cell- or tissue-specific secreted and surface proteins, a polyclonal antiserum was raised against a complex mixture of surface or secreted proteins from the target cell, followed by immunodepletion of antibodies that recognize proteins from a nontarget cell or...

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Bibliographic Details
Published in:Nature biotechnology 1998-06, Vol.16 (6), p.581-586
Main Authors: Scherer, Philipp E., Bickel, Perry E., Kotler, Mariana, Lodish, Harvey F.
Format: Article
Language:English
Subjects:
3T3 Cells
Adipocytes - chemistry
Adipocytes - immunology
Adipocytes - secretion
Agriculture
Amino Acid Sequence
Animals
Antibody Specificity
Bioinformatics
Biological and medical sciences
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
Cell Line
Chemical Precipitation
Cloning, Molecular
Collagen - genetics
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Immune Sera - chemistry
Immune Sera - isolation & purification
Immune Sera - metabolism
Life Sciences
Membrane Proteins - immunology
Membrane Proteins - secretion
Methods. Procedures. Technologies
Mice
Molecular cloning
Molecular Sequence Data
Reproducibility of Results
research-article
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Summary:To identify and clone genes that encode cell- or tissue-specific secreted and surface proteins, a polyclonal antiserum was raised against a complex mixture of surface or secreted proteins from the target cell, followed by immunodepletion of antibodies that recognize proteins from a nontarget cell or tissue. The depleted antiserum is used to screen bacteriophage cDNA expression libraries. Because of our interest in how adipocytes communicate with other cells, we have used this method to clone cDNAs encoding secreted and plasma membrane proteins that are induced during adipocyte differentiation. We describe several of these, including a novel plasma membrane-associated protein, S3-12.
ISSN:1087-0156
1546-1696
DOI:10.1038/nbt0698-581