Cell lysates prepared in RIPA with 1X HALT. RNaseOUT (Thermo Fisher Scientific) RNase inhibitor was added to the lysis buffer at 0.5 U/µL when RNase A was not used. A total of 1 mg of protein lysate was mixed with 2–10 µg of IgG or specific antibody overnight at 4°C with rotation. For samples treated with RNase A, 20 µg of RNase A (Invitrogen) or 20 U RNase III with MnCl2 added to 20 nmol/L (NEB), was added to the lysate during overnight mixing with the antibody. Protein G Dynabeads (Thermo Fisher Scientific, 25 µL per sample) were prepared by washing twice in the lysis buffer. Prepared beads were mixed with lysates for 30 minutes at 4°C with rotation. Supernatants were collected and beads were washed three times in the lysis buffer, and eluted by mixing the beads in SDS sample buffer and incubating at 95°C for 7 minutes. Antibodies: Rabbit IgG (Jackson ImmunoResearch, 011-000-003), Mouse IgG (Jackson ImmunoResearch Labs, catalog no. 015-000-003, RRID:AB_2337188), DHX9 (Bethyl, catalog no. A300-855A, RRID:AB_609442), PARP (Cell Signaling Technology, catalog no. 9532, RRID:AB_659884), XRN2 (Novus, catalog no. NB100-57541, RRID:AB_2288770), DDX54 (Novus, catalog no. NB100-60678, RRID:AB_921120), DDX17 (Thermo Fisher Scientific, catalog no. PA5-84585, RRID:AB_2791736).
Ddx17
Manufactured by Thermo Fisher Scientific
The DDX17 is a lab equipment product from Thermo Fisher Scientific. It is a protein that functions as an RNA helicase, which is an enzyme that unwinds double-stranded RNA molecules. The DDX17 protein is involved in various cellular processes, including transcription, splicing, and ribosome biogenesis.
Automatically generated - may contain errors
2 protocols using ddx17
1
RNA-binding Protein Immunoprecipitation Protocol
2
Immunoprecipitation of RNA-binding proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cell lysates prepared in RIPA with 1X HALT. RNaseOUT (Thermo Fisher) RNase inhibitor was added to the lysis buffer at 0.5 U/μL when RNase A was not used. One milligram of protein lysate was mixed with 2–10 μg of IgG or specific antibody overnight at 4 °C with rotation. For samples treated with RNase A, 20 μg of RNase A (Invitrogen) was added to the lysate during overnight mixing with the antibody. Protein G Dynabeads (Thermo Fisher, 25 μL per sample) were prepared by washing twice in the lysis buffer. Prepared beads were mixed with lysates for 30 min at 4 °C with rotation. Supernatants were collected and beads were washed three times in the lysis buffer, and eluted by mixing the beads in SDS sample buffer and incubating at 95 °C for 7 min. Antibodies: Rabbit IgG (Jackson ImmunoResearch, 011-000-003), Mouse IgG (Jackson ImmunoResearch, 015-000-003), DHX9 (Bethyl, A300-855A), PARP (Cell Signaling, 9532S), XRN2 (Novus, NB100-57541), DDX54 (Novus, NB100-60678), DDX17 (Thermo Scientific, PA5-84585).
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