Comprehensive Antibody Immunodetection Protocol

Antibodies recognizing the Flag tag (Sigma,F1804), SETX (Novus, NBP1-94712), RNA polymerase I (RPA194 (C-1) (Santa Cruz, sc-48385), fibrillarin (G-8) (Santa Cruz, sc-374022), nucleophosmin (NPM1, Forties Life Sciences A302-403A-T), C1QBP/P32 (Proteintech, 24474-1-AP), CHCHD2 (Proteintech, 66302-1-lg), GAPDH (Sigma-Aldrich, G9545), U2AF65 (Sigma-Aldrich, U4758), H3 (Abcam, ab8898), phospho-histone H2A.X(Ser139) (Cell signaling, 2577), and MitoTracker Red CMXRos (Invitrogen, M7512) were obtained from the indicated vendors. Human RIEP rabbit polyclonal antibodies were generated at Bethyl Laboratories/Fortis Life Sciences. RIEP-1 and RIEP-216 antibodies recognize peptides derived from the RIEP N terminus or C terminus, respectively. IGPAL-CA-630 (Sigma, I8896), cOmplete™, Mini, EDTA-free Protease Inhibitor Cocktail (Roche, 04693159001), and Pierce™ Protein A/G Magnetic Beads (Thermo Fisher scientific, 88802) were purchased from indicated vendors.

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Feng S., Desotell A., Ross A., Jovanovic M, & Manley J.L. (2023). A nucleolar long “non-coding” RNA encodes a novel protein that functions in response to stress. Proceedings of the National Academy of Sciences of the United States of America, 120(9), e2221109120.

Publication 2023

F1804 RPA194 (C-1) fibrillarin (G-8) GAPDH U2AF65 phospho-histone H2A.X(Ser139) MitoTracker Red CMXRos IGPAL-CA-630 EDTA-free Protease Inhibitor Cocktail Pierce™ Protein A/G Magnetic Beads

Antibodies Edta Fibrillarin Gapdh Histone h2a x Human Mitotracker red cmxros Novus Npm1 Peptides Protease inhibitor Protein a Protein g Rabbit Rna polymerase i Setx

Corresponding Organization :

Other organizations : Columbia University

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Variable analysis

independent variables

Antibodies recognizing the Flag tag (Sigma,F1804)
SETX (Novus, NBP1-94712)
RNA polymerase I (RPA194 (C-1) (Santa Cruz, sc-48385)
Fibrillarin (G-8) (Santa Cruz, sc-374022)
Nucleophosmin (NPM1, Forties Life Sciences A302-403A-T)
C1QBP/P32 (Proteintech, 24474-1-AP)
CHCHD2 (Proteintech, 66302-1-lg)
GAPDH (Sigma-Aldrich, G9545)
U2AF65 (Sigma-Aldrich, U4758)
H3 (Abcam, ab8898)
Phospho-histone H2A.X(Ser139) (Cell signaling, 2577)
MitoTracker Red CMXRos (Invitrogen, M7512)
RIEP-1 and RIEP-216 antibodies

dependent variables

Not explicitly mentioned

control variables

IGPAL-CA-630 (Sigma, I8896)
COmplete™, Mini, EDTA-free Protease Inhibitor Cocktail (Roche, 04693159001)
Pierce™ Protein A/G Magnetic Beads (Thermo Fisher scientific, 88802)

positive controls

Not specified

negative controls

Not specified

Annotations

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