BCAR1Breast Cancer Antiestrogen Resistance 1
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Additional protein preparations relating to BCAR1 were used in Western blotting experiments and in assay specificity studies.
We estimated the amounts of BCAR1 obtained by measuring the absorbance of eluted fractions at 280 run.
Polyclonal antibodies against BCAR1, isolated from chicken egg yolks and citrate plasma from rabbits, were purified by affinity chromatography on GST-BCAR1/F1R5-coated columns (AffiGel[R]15; Bio-Rad Laboratories) according to previously described procedures (8,10).
The polyclonal antibodies against BCAR1, raised in chickens (328#) and rabbits (329#), were used diluted 300- to 10 000-fold.
BCAR1 concentrations were measured by ELISA with the same experimental setup as described previously by Grebenschikov et al.
Purified His-BCAR1/F1R5 was used as a calibrator in the BCAR1 ELISA to generate the dose-response curve.
Assay specificity was checked by analyzing different dilutions of four cytosol preparations based on ZR-75-1 breast cancer cell-derived stable transfectants (ZR-75-1, F-BCAR1, BCAR1, and F-Hybrid).
To develop antibodies specific for BCAR1, we selected a central part of the BCAR1 cDNA that encoded residues 272-541 of the protein (Fig.
To analyze the relationship of BCAR1 with benefit of tamoxifen therapy, the 592 patients were divided into four equal-sized groups based on the quartile protein concentrations of BCAR1.
For the BCAR1 concentrations treated as log-transformed continuous variables [ln([micro]g/g of protein)], the multivariable OR was 0.68 (95% confidence interval, 0.47-0.99; P = 0.044).
This study shows that BCAR1 protein concentrations in the primary tumor can predict the efficacy of tamoxifen therapy in recurrent breast cancer.
Development of an ELISA for measurement of BCAR1 protein in human breast cancer tissue.