1. Evaluation of Antibody Diluents for FANCD2 Immunohistochemistry with Breast Cancer Tissue
- Author
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Stella Quan, Xuelin Cui, Sharron Ladd, Brian E. Ward, and Hua Chang
- Subjects
Tris ,Pathology ,medicine.medical_specialty ,Histology ,Chromatography ,biology ,Chemistry ,medicine.disease ,Isotype ,Diluent ,Staining ,Medical Laboratory Technology ,chemistry.chemical_compound ,fluids and secretions ,Breast cancer ,Casein ,biology.protein ,medicine ,Immunohistochemistry ,Anatomy ,Antibody - Abstract
Two important criteria used to select an antibody diluent for robust immunohistochemistry are: (1) optimal signal/noise ratio, and (2) long-term stability when diluted. For immunohistochemistry with Fanconi anemia, complementation group D2 (FANCD2) in formalin-fixed-paraffin-embedded breast cancer tissue, eight experimental and eight commercial diluents were evaluated. Staining intensity was quantitated with Aperio digital image analysis software. An H-Score was calculated for FANCD2 and isotype control staining for each diluent. Signal/noise ratio was defined as H-score of the antibody divided by H-score of the isotype control. The ratios ranged from 0.46 to 135.9 for different diluents. Casein in Tris-based buffer produced the greatest signal/noise ratio. The two best commercial and four best experimental diluents were selected for evaluation of the diluted FANCD2 antibody stability at 4°C, room temperature, and 37°C, respectively. Three diluents were eliminated from further investigation after ...
- Published
- 2010