Development of an ELISA for Measurement of BCAR1 Protein in Human Breast Cancer Tissue

High concentrations of breast cancer anti-estrogen resistance 1 (BCAR1) protein measured by Western blotting in primary breast tumor cytosols are associated with early disease progression and failure of tamoxifen therapy. The aim of the present study was to develop an ELISA to measure BCAR1 quantita...

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Bibliographic Details
Published in:Clinical chemistry (Baltimore, Md.) Md.), 2004-08, Vol.50 (8), p.1356-1363
Main Authors: Grebenchtchikov, Nicolai, Brinkman, Arend, van Broekhoven, Simone P.J, de Jong, Danielle, Geurts-Moespot, Anneke, Span, Paul N, Peters, Harry A, Portengen, Henk, Foekens, John A, Sweep, C.G.J, Dorssers, Lambert C.J
Format: Article
Language:English
Subjects:
Adapter proteins
Amino acids
Animals
Antibodies
Antibody Specificity
Blotting, Western
Breast cancer
Breast Neoplasms - chemistry
Breast Neoplasms - diagnosis
Cancer therapies
Cell Line, Tumor
Chickens
Crk-Associated Substrate Protein
Disease
Enzyme-Linked Immunosorbent Assay - methods
Estrogens
Female
Humans
Predictive Value of Tests
Prognosis
Proteins - analysis
Rabbits
Retinoblastoma-Like Protein p130
RNA polymerase
Sensitivity and Specificity
Tumors
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Summary:High concentrations of breast cancer anti-estrogen resistance 1 (BCAR1) protein measured by Western blotting in primary breast tumor cytosols are associated with early disease progression and failure of tamoxifen therapy. The aim of the present study was to develop an ELISA to measure BCAR1 quantitatively in extracts of human breast cancer tissue. A recombinant fragment of BCAR1 (the human homolog of murine p130Cas) was produced in bacterial M15 cells, purified, and injected into chickens and rabbits. The generated antibodies were affinity-purified and used for the construction of an ELISA. After validation, the results obtained with the ELISA were compared with Western blot findings on primary breast tumors. The detection limit the BCAR1 ELISA was 0.0031 microg/L, and the within-run imprecision (CV) was
ISSN:0009-9147
1530-8561
DOI:10.1373/clinchem.2003.029868