National Toxicology Program

Tables of Contents: In Vitro Endocrine Disruptor Final Background Review Documents (BRDs)
October 2002

• Estrogen Receptor Binding


• Estrogen Receptor Transcriptional Activation


• Androgen Receptor Binding


• Androgen Receptor Transcriptional Activation

• Final ER Binding BRD - NIH Publication No. 03-4504 in PDF [5.7 MB]


• Appendix A - Methods for In Vitro ER Binding Assays
• A1 - Assays Using Rat Uterine Cytosol
• A2 - Assays Using Mouse Uterine Cytosol
• A3 - Assays Using Human ER a and ERb
• A4 - Assays Using Recombinant ER a and ERb
• A5 - Assays Using Fluorescence Polarization
• A6 - Assays Using GST-ERdef Constructs
• A7 - Assays Using MCF-7 Cells and Cytosol


• Appendix B - In Vitro ER Binding Assay Protocols
• B1 - Protocol for the Competitive ER binding MCF-7 (Whole Cell Assay)
• B2 - Protocol for the Fluorescence Polarization Assay of the Competitive Binding of Ligands to Estrogen-Receptor Complexes
• B3 - Protocol for the Competitive Ligand Binding Assay
• B4 - Standard Operating Procedure for the Rat Estrogen Receptor Equilibrium Exchange Assay
• B5 - Protocol for the Estrogen Receptor Competitive Binding Assay Using Rat Uterine Cytosol


• Appendix C - Chemical and Product Class Information for the Substances Tested in the In Vitro ER Binding Assays


• Appendix D - Substances Tested in the In Vitro ER Binding Assays
• D1 - Data Sorted by Substance Name and Assay
• D2 - References


• Appendix E - Assay Distribution of Substances Tested in In Vitro ER Binding Assays


• Appendix F - Median/Single RBA Values for Substances Tested in Two or More In Vitro ER Binding Assays

• Final ER TA BRD - NIH Publication No. 03-4505 in PDF [4.8 MB]


• Appendix A - Methods for In Vitro ER TA Assays
• A1 - ER TA Reporter Gene Assays Using MCF-7 Cells
• A2 - ER TA Reporter Gene Assays Using Various Mammalian Cells
• A3 - ER TA Cell Proliferation Assays
• A4 - ER TA Reporter Gene Assays Using Yeast Cells


• Appendix B - In Vitro ER TA Assay Protocols
• B1 - Protocol for HepG2 Cells + Receptor + Reporter and/or b-gal Plasmids for Use in Steroid Hormone Receptor Assays
• B2 - Protocol for Chimeric ERa-Mediated Reporter Gene Expression in MCF-7 Cells
• B3 - Protocol for the Development of new reporter gene assay systems for screening Endocrine Disruptors
• B4 - Protocol for the Development of stably transfected cell lines to screen Endocrine Disruptors
• B5 - Technical Perspective on the U.S.E.P.A. Endocrine Disruptor Screening Program: In Vitro EDSTAC Guideline Protocols
• B6 - Lyticase-based cell lysis protocol of b-Galactosidase assay for 96 well plates
• B7 - Protocol for High-Throughput System for Screening Estrogen-Like Chemicals
• B8 - Protocol for the MVLN Assay


• Appendix C - Chemical and Product Class Information for Substances Tested in ER TA Assays


• Appendix D - Substances Tested in the In Vitro ER TA Assays
• D1 - Data Sorted by Chemical Name and Assay
• D2 - References


• Appendix E - Responses of Substances Tested in at Least Two Different Types of In Vitro ER TA Assays


• Appendix F - Substances Tested in Different In Vitro ER TA Assays by the Same Investigator

• Final AR Binding BRD - NIH Publication No. 03-4506 in PDF [3.8 MB]


• Appendix A - Methods for In Vitro AR Binding Assays
• A1 - Assays Using Rat Prostate or Epididymal Cytosol
• A2 - Assays Using Human Genital Fibroblast Cells
• A3 - Assays Using COS Cells Transfected with AR
• A4 - Assays Using MCF-7 Cells
• A5 - Miscellaneous AR Binding Assays


• Appendix B - In Vitro AR Binding Assay Protocols
• B1 - Protocol for Androgen Receptor Competitive Binding Assay Using Rat Prostate Cytosol
• B2 - Protocol for COS Cell Binding Assay
• B3 - Protocol for Measuring Androgen-Binding Sites on Androgen Receptors or Binding Proteins
• B4 - Technical Perspective on the U.S.E.P.A. Endocrine Disruptor Screening Program: In Vitro EDSTAC Guideline Protocols


• Appendix C - Chemical and Product Class Information for the Substances Tested in the In Vitro AR Binding Assays


• Appendix D - Substances Tested in the In Vitro AR Binding Assays
• D1 - Sorted by Substance Name, Assay and Reference Androgen
• D2 - References

• Final AR TA BRD - NIH Publication No. 03-4507 in PDF [4.4 MB ]



• Appendix A - Methods for In Vitro AR TA Assays
• A1 - AR TA Assays Using CHO Cells
• A2 - AR TA Assays Using Human Cells
• A3 - AR TA Assays Using CV-1 Cells
• A4 - AR TA Assays Using Yeast Cells
• A5 - Miscellaneous AR TA Assays


• Appendix B - In Vitro AR TA Assay Protocols
• B1 - Protocol for CV1 + hAR + Luciferase Assay
• B2 - Protocol for CHO Cells + hAR + Luciferase Assay
• B3 - Protocol for HepG2 Cells + Receptor + Reporter and/or b-gal Plasmids for Use in Steroid Hormone Receptor Assays
• B4 - Protocol for Yeast-Based Androgen Receptor Assay
• B5 - Protocol for the Development of new reporter gene assay systems for screening Endocrine Disruptors
• B6 - Protocol for the Development of stably transfected cell lines to screen Endocrine Disruptors
• B7 - Technical Perspective on the U.S.E.P.A. Endocrine Disruptor Screening Program: In Vitro EDSTAC Guideline Protocols


• Appendix C - Chemical and Product Class Information for Substances Tested in AR TA Assays


• Appendix D - Data for Substances Tested in the In Vitro AR TA Assays
• D1 - Sorted by Chemical Name and Assay
• D2 - References


• Appendix E - Substances Tested in Two or More Selected In Vitro AR TA Assays
• E1 - Substances Tested for Agonism Activity
• E2 - Substances Tested for Antagonism Activity