Monoclonal Antibodies Against BAP1 (BRCA1-Associated Proteins)

Inventor(s)
Tech ID
RF96001
Background

Wistar researchers have identified a novel protein, BAP1, which is a nuclear-localized, ubiquitin carboxyl-terminal hydrolase that binds to the wild-type RING finger domain of the Breast/Ovarian Cancer Susceptibility Gene product, BRCA1. Murine Bap1 and Brca1 are temporally and spatially co-expressed during murine breast development and remodeling, and show overlapping patterns of subnuclear distribution. BAP1 resides on human chromosome 3p21.3 and rearrangements, deletions and missense mutations of BAP1 have been found in lung carcinoma cell lines and in primary breast tumor samples. BAP1 enhances BRCA1-mediated inhibition of breast cancer cell growth and is the first nuclear-localized ubiquitin carboxy-terminal hydrolase to be identified.

Origin

BAP1 mAbs are secreted by hybridomas derived from mice immunized with full-length BAP1. This protein was expressed and purified from E.coli. Individual mAbs were selected for reactivity to various domains of BAP1 fused to GST. Mab 1G8 and Mab 2A2 recognize epitopes within the UCH domain of BAP1. Mab 3C11 recognizes an epitope within the region of amino acids 337-440.

Reactivity

All mAbs recognize full-length BAP1 produced by 1) in vitro transcription/translation; 2) over-expression in transiently transfected COS1 cells; and 3) over-expression from stably transfected MCF7 cells {breast cancer cell line} and from stably transfected NCI-H226 cells (non-small cell lung carcinoma cell line).

Isotope
All are IgG1k
Applications and Advantages

These antibodies have been used in immunoprecipitation and immunofluorescense analyses.