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Literature DB >> 9254718

A solid-phase assay to screen monoclonal antibodies against DNA-binding protein.

N Ishii¹, H Nakayama, J Katayama, M Arisawa, Y Aoki.

Abstract

A method is described for selecting monoclonal antibodies (mAb) against DNA-binding protein. The protocol involves a non-radioactive solid-phase DNA binding assay using a 96-well plate. Because the solid-phase assay is highly specific and sensitive, partially purified antigen is sufficient for the immunization, and mAb screening can be performed with crude cell extract as the antigen. MAbs obtained by this method could supershift the DNA-protein complex in the electromobility shift assay, and were sufficient for immunoscreening of a cDNA expression library.

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Year: 1997 PMID： 9254718 PMCID： PMC146919 DOI： 10.1093/nar/25.17.3550

Source DB: PubMed Journal: Nucleic Acids Res ISSN： 0305-1048 Impact factor: 16.971

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References

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7 in total

A solid-phase assay to screen monoclonal antibodies against DNA-binding protein.

1. Colorimetric detection of specific DNA segments amplified by polymerase chain reactions.

2. A DNA-binding protein from Candida albicans that binds to the RPG box of Saccharomyces cerevisiae and the telomeric repeat sequence of C. albicans.

3. Molecular cloning of an enhancer binding protein: isolation by screening of an expression library with a recognition site DNA.

4. Purification and cloning of a DNA binding protein from yeast that binds to both silencer and activator elements.

5. Isolation of ORC6, a component of the yeast origin recognition complex by a one-hybrid system.

6. Zinc finger phage: affinity selection of fingers with new DNA-binding specificities.

7. Purification and characterization of a nuclear factor which binds specifically to the upstream activation sequence of Saccharomyces cerevisiae enolase 1 gene.