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BID binds USP9X. 13 / 13
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"We then obtained a recombinant BFL-1 with C55A mutation (BFL-1 C55A ) and determined that BFL-1 C55A had an IC 50 value of 5.41 nM with the FAM-BID 79-99 tracer in the titration assay (SI, Fig. S5 )."
35385805
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"Dissociation constant ( K d ) of the FAM-BID was determined to be 5.99 ± 1.12 nM, demonstrating stable signal up to 24h ( Fig. S3 )."
37562606
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"Based on the K d value, the concentration of the Mcl-1 protein used in the competitive binding experiments was 20 nM and the fluorescent probe, FAM-BID, was fixed at 2 nM."
37562606
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"The fluorescein tagged BID BH3 peptide (79–99), named as FAM-BID, was synthesized by the Synpeptide.com (China)."
35385805
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"Their K d values to BFL-1 (1–151)/FAM-BID and BFL-1 C55A (1–151)/FAM-BID were determined with a fixed concentration of FAM-BID and a serial concentration of BFL-1 or BFL-1 C55A protein in saturation e[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
35385805
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"Then, 20 μL FAM-BID peptide in PBS was added to the protein-compound mixture solution and incubated at 37 °C for 20 min."
38947842
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"To each single well, 2 components were added sequentially: (1) 4 μL of the serial diluted testing compounds in DMSO; (2) The FAM-BID was added to the assay buffer comprising 100 mM potassium phosphate[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
35385805
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"A 21-residue Bid BH3 peptide (residues 79–99:QEDIIRNIARHLAQVGDSMDR) derived from its BH3 domain, was synthesized and labeled at the N-terminus with 6-carboxyfluorescein succinimidyl ester (FAM) as the[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
22739087
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"Saturation experiments determined that FAM-Bid binds to Bcl-2 protein with a K d value of 7.1 nM."
22739087
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"For competitive binding experiments, Bcl-2 protein (40 nM) and FAM-Bid peptide (2.5 nM) were preincubated in the assay buffer (100 mM potassium phosphate, pH 7.5; 100 μg/mL bovine gamma globulin; 0.02[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
22739087
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"5 μL of a solution in DMSO of the tested compound was added to the Bcl-2/FAM-Bid solution in Dynex 96-well, black, round-bottom plates (Fisher Scientific) to produce a final volume of 125 μL. For each[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
22739087
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"In order to screen compounds that competitively block the BFL-1/BH3s interactions, a fluorescent polarization (FP) assay with an IC 50 value of 3.31 nM was established based on a FAM labeled BID pepti[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
35385805
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"Then, 20 μL of a solution in phosphate buffered saline (PBS) of the FAM-Bid peptide was added to the solution to produce a final volume of 200 μL and incubated at 37 °C for 20 min."
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