Identification of microinjected cells using biotinylated antibodies and Strep-avidin-conjugated horseradish peroxidase

Riabowol, K. T. (1988) Identification of microinjected cells using biotinylated antibodies and Strep-avidin-conjugated horseradish peroxidase. Anal Biochem, 174 (2). pp. 601-12. ISSN 0003-2697

URL: http://www.ncbi.nlm.nih.gov/pubmed/3239762
DOI: 10.1016/0003-2697(88)90062-0

Abstract

Results from experiments using needle microinjection of cells are often compromised by an inability to readily demonstrate which cells within a population have been injected. The technique described here allows the unambiguous identification of cells that have been successfully microinjected. Sequential incubation of fixed cells with biotinylated anti-immunoglobulin antibodies, followed by horseradish peroxidase (HRP)-conjugated Strep-avidin and HRP substrate, provides a sensitive assay for identification of cells containing trace amounts of immunoglobulins. This allows direct correlation to the presence of injected molecules of effects on cell morphology, the ability to enter into DNA synthesis, or expression of specific genes. By a variety of criteria, nonspecific immunoglobulins do not adversely affect cellular processes when injected by themselves or in the presence of other proteins known to have biological effects when injected, such as cAMP-dependent protein kinase and the ras oncogene protein.

Item Type: Paper
Subjects: Investigative techniques and equipment
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > antibodies
CSHL Authors:
Communities: CSHL labs
Depositing User: Gail Sherman
Date Deposited: 19 Sep 2017 19:34
Last Modified: 19 Sep 2017 19:34
Related URLs:
URI: http://repository.cshl.edu/id/eprint/35203

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