Comparative Analysis of Mitosis-Specific Antibodies For Bulk Purification of Mitotic Populations By Fluorescence-Activated Cell Sorting | Zendy

Amy E. Campbell | Zendy; Chris C.S. Hsiung | Zendy; Gerd A. Blobel | Zendy

Open Access

Comparative Analysis of Mitosis-Specific Antibodies For Bulk Purification of Mitotic Populations By Fluorescence-Activated Cell Sorting

Author(s) -

Amy E. Campbell,

Chris C.S. Hsiung,

Gerd A. Blobel

Publication year - 2014

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/000114137

Subject(s) - mitosis , interphase , cell sorting , biology , microbiology and biotechnology , epitope , chromatin , genome , computational biology , cell , antibody , genetics , dna , gene

Mitosis entails complex chromatin changes that have garnered increasing interest from biologists who study genome structure and regulation-fields that are being advanced by high-throughput sequencing (Seq) technologies. The application of these technologies to study the mitotic genome requires large numbers of highly pure mitotic cells, with minimal contamination from interphase cells, to ensure accurate measurement of phenomena specific to mitosis. Here, we optimized a fluorescence-activated cell sorting (FACS)-based method for isolating formaldehyde-fixed mitotic cells--at virtually 100% mitotic purity and in quantities sufficient for high-throughput genomic studies. We compared several commercially available antibodies that react with mitosis-specific epitopes over a range of concentrations and cell numbers, finding antibody MPM2 to be the most robust and cost-effective.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research