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 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.
Scherer
P E
PE
Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.
Bickel
P E
PE
Kotler
M
M
Lodish
H F
HF
D016475:3T3 Cells; D017667:Adipocytes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D002460:Cell Line; D011232:Chemical Precipitation; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics; D007106:Immune Sera / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008565:Membrane Proteins / Q000276:immunology* / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D015203:Reproducibility of Results
D007106:Immune Sera; D008565:Membrane Proteins; D003094:Collagen