Download The Nuclear Receptor Superfamily: Methods and Protocols by Iain J. McEwan (auth.), Iain J. McEwan (eds.) PDF
By Iain J. McEwan (auth.), Iain J. McEwan (eds.)
Acting largely to manage styles of gene expression, nuclear receptors play important roles in the course of embryonic improvement and within the law of metabolic and reproductive capabilities in grownup existence, which proves this superfamily of ligand-activated transcription components to be an important a part of organic existence. In The Nuclear Receptor Superfamily: equipment and Protocols, specialist researchers describe a variety of molecular, structural and phone organic concepts at present used to enquire the structure-function of nuclear receptors, including experimental techniques that could result in new healing suggestions for treating nuclear receptor-associated illnesses. Written within the hugely profitable Methods in Molecular Biology™ sequence structure, the chapters during this quantity comprise short introductions to the themes, lists of the mandatory fabrics and reagents, step by step, easily reproducible laboratory protocols, in addition to notes from the specialists to spotlight pointers on troubleshooting and warding off recognized pitfalls.
Cutting-edge and straightforward to exploit, The Nuclear Receptor Superfamily: tools and Protocols offers important and time-saving assistance for all these venture learn during this ever-growing box of study.
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2% sodium azide. 3. 1 mg/mL; BD Biosciences; San Diego, CA). 1 mg/mL) (BD Biosciences). 38 Butts and Sternberg 4. Secondary Antibodies: fluorescein isothyocyanate (FITC)conjugated goat anti-rabbit antibody (for GR and AR); fluorescein isothyocyanate (FITC)-conjugated goat anti-mouse antibody (for PR). 5. Cytofix/Cytoperm solution and wash buffer (BD Biosciences, San Diego, CA). 6. Rat serum (Sigma) to prevent nonspecific binding of antibodies within the cytoplasm of cells. 7. Paraformaldehyde solution (4%) to generate fixed cells if cells cannot be examined within 24–48 h of collection and staining with antibodies.
Appropriate safety measures and adherence to local and national rules should be followed when working with ionizing radiation sources and discarding waste materials. 625 mL and add unlabeled ligand to 500 nM (equivalent to a 100-fold excess of Methods for Measuring Ligand Dissociation and Nuclear Receptor Turnover 27 [3H]-ligand). 1 mL of the hot and hot + cold labeling solutions to determine the total radioactivity in a scintillation counter. 2. Label the plates according to the selected time points.
8. Butts CL, Shukair SA, Duncan KM, et al. Progesterone inhibits mature rat dendritic cells in a receptor-mediated fashion. Int Immunol 2007;19(3):287–96. 9. Butts CL, Shukair SA, Duncan KM, Harris CW, Belyavskaya E, Sternberg EM. Effects of dexamethasone on rat dendritic cell function. Horm Metab Res 2007;39(6):404–12. 10. Butts CL, Shukair SA, Duncan KM, Harris CW, Belyavskaya E, Sternberg EM. Evaluation of steroid hormone receptor protein expression in intact cells using flow cytometry. Nucl Recept Signal 2007;5:e007.