Production and characterization of monoclonal antibodies to the steroid receptor coactivator AIB1.
The steroid coactivator AIB1 (Ampified In Breast cancer 1) is a member of the SRC-1 family of transactivation coactivators that is amplified in about 7% of breast tumor samples. Hybridomas were established that secreted monoclonal antibodies (MAbs) to a recombinant glutathione-S-transferase (GST) fusion protein expressing the steroid receptor interacting domain of AIB1 (GST-AIB.T1:a.a. 605-1294). Four MAbs from these hybridomas were characterized and designated AX15.1, AX15.2, AX15.3, and AX15.4. The MAbs were shown to be specific to AIB1 and did not cross-react with two similar coactivators SRC-1 and TIF2 as shown in Western blot analysis and enzyme-linked immunoadsorbent assay (ELISA). Western blot analysis using the four MAbs showed specific recognition of AIB1 protein as a 160 kDa band in lysates from cell lines containing AIB1 gene-amplification. The MAbs immunoprecipitated in vitro-translated AIB1 and cellular AIB1 from metabolically labeled cells. The results show that these newly described MAbs are useful in studies of AIB1 function and expression.
Azorsa
D O
DO
Cancer Genetics Branch, National Human Genome Research Institute, The National Institutes of Health, Bethesda, MD 20892, USA.
Meltzer
P S
PS
D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D051548:Histone Acetyltransferases; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D056920:Nuclear Receptor Coactivator 1; D011233:Precipitin Tests; D011987:Receptors, Steroid / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D015534:Trans-Activators / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D014157:Transcription Factors / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D011987:Receptors, Steroid; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D051548:Histone Acetyltransferases; C538859:NCOA1 protein, human; D056920:Nuclear Receptor Coactivator 1