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MR-LBD™ | Cat. #S28 |
Purified GST-tagged human Mineralocorticoid receptor LBD/Hsp90 complex |
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GST-tagged Mineralocorticoid receptor ligand-binding domain (MR-LBD) consists of a 218 a.a. GST tag (Schistosoma japonicum Glutathione S-Transferase, Acc# 1M99_A), a 30 a.a. flexible linker and a 381 a.a. MR-LBD, Acc# AAA59571, amino acids 604-984 (details). Complex of MR with human Hsp90 molecular chaperone is necessary for attaining native ligand-binding competent conformation and receptor activity in ligand binding and coactivator peptide recruitment assays (details). To this effect a complex of human MR-LBD with human Hsp90 (MR-LBD/Hsp90) was produced in insect cells co-infected with two recombinant baculoviruses, i.e. BV-MR-LBD™ that encodes GST-tagged MR-LBD and FoldHelper™-905c which provides for expression of Hsp40, Hsc70, Hsp90, Hop and p23 human molecular chaperones. Cells were cultivated in the absence of exogeneous ligands to ensure availability of the ligand-binding site. MR-LBD/Hsp90 complex was purified from soluble cell fraction without the use of detergents, high-salt buffers or chaotropic agents and was formulated at about physiological conditions. GST tagged MR-LBD and Hsp90 are the major components of the complex (Fig. 1). |
Storage buffer 20 mM Na-P pH 8.0, 40 mM KCl, 20 mM Na2MoO4, 7 mM reduced glutathione, 15% glycerol. Purity >90% Applications Ligand binding and coactivator peptide recruitment assays. Handling upon arrival Immediately store at -80°C. Avoid freeze-thaw cycles. While working, please keep sample on ice. |
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Fig. 1. SDS-PAGE analysis of purified human Mineralocorticoid receptor LBD in complex with Hsp90. Proteins were separated in 4-20% SDS-PAGE and stained with Coomassie blue. Identity of MR-LBD, Hsp90 and Hsc70 was confirmed using mass-spectrorometry. Open the links above for amino acid sequence coverage. 1. Mineralocorticoid receptor LBD/Hsp90 complex. 2. Mark12™ Unstained Standard (Life Technologies). |