Antibody Characterization for Protein Detection

The following antibodies were used for immunofluorescence (IF) and immunoblotting (IB) at indicated dilutions: sheep anti-pan-RNase H2 (raised against human recombinant RNase H213 (link), IB 1:1,000, IP 5 µl / 1 ml lysate); rabbit anti-RNASEH2C (Proteintech 16518-1-AP; IB 1:1,000); rabbit anti-RNASEH2A (Origene TA306706, IB 1:1,000); mouse anti-RNASEH2A (Abcam ab92876; IF 1:500); mouse anti-RNASEH2A G-10 (Santa Cruz Biotechnologies sc-515475; WB 1:1000); mouse anti-γH2AX JBW301 (Millipore 05-636, IF 1:800 – 1:2,000); rabbit anti-RAD51 H-92 (Santa Cruz Biotechnologies sc-8349, IF 1:150); rabbit anti-BRCA146 (link) (IB 1:1,000); mouse anti-Cas9 (Diagenode C15200203, IB 1:1,000); rabbit anti-PARP1 H-250 (Santa Cruz Biotechnologies sc-7150, IB 1:1,000); mouse anti-PAR 10H (Enzo ALX-804-220-R100, IB 1:1,000); rabbit anti-Topoisomerase I (Abcam ab109374; IB 1:5,000); rabbit anti-DYKDDDDK (Cell Signaling Technologies 2368, IB 1:1,000); rabbit anti-actin (Sigma A2066, IB 1:5,000); mouse anti-α–tubulin DM1A (Millipore CP06, IB 1:5,000); mouse anti-α–tubulin B512 (Sigma T6074, IB 1:5,000); rabbit anti-GAPDH (Sigma G9545, IB 1:20,000); mouse anti-vinculin (Sigma V9264, IB 1:1,000); rabbit anti-DNA polymerase beta (Abcam ab26343, IB 1:1,000); rabbit anti-cleaved caspase-3 (Cell Signaling Technologies 9661S, IF 1:800).

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Zimmermann M., Murina O., Reijns M.A., Agathanggelou A., Challis R., Tarnauskaitė Ž., Muir M., Fluteau A., Aregger M., McEwan A., Yuan W., Clarke M., Lambros M.B., Paneesha S., Moss P., Chandrashekhar M., Angers S., Moffat J., Brunton V.G., Hart T., de Bono J., Stankovic T., Jackson A.P, & Durocher D. (2018). CRISPR screens identify genomic ribonucleotides as a source of PARP-trapping lesions. Nature, 559(7713), 285-289.

Publication 2018

rabbit anti-RNASEH2C rabbit anti-RNASEH2A mouse anti-RNASEH2A mouse anti-RNASEH2A G-10 mouse anti-γH2AX JBW301 rabbit anti-RAD51 H-92 mouse anti-Cas9 rabbit anti-PARP1 H-250 mouse anti-PAR 10H rabbit anti-Topoisomerase I rabbit anti-DYKDDDDK rabbit anti-actin mouse anti-α–tubulin DM1A mouse anti-α–tubulin B512 rabbit anti-GAPDH mouse anti-vinculin rabbit anti-DNA polymerase beta rabbit anti-cleaved caspase-3

Actin Anti topoisomerase i Antibodies Caspase 3 Dilutions Dna polymerase beta Gapdh Human Immunofluorescence Mouse Parp1 Rabbit Rnase Sheep Vinculin α tubulin

Corresponding Organization :

Other organizations : Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, University of Birmingham, Edinburgh Cancer Research, University of Toronto, Institute of Cancer Research, Heartlands Hospital, The University of Texas MD Anderson Cancer Center

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Variable analysis

independent variables

Antibodies used for immunofluorescence (IF) and immunoblotting (IB)

dependent variables

Immunofluorescence (IF) staining
Immunoblotting (IB) or Western blotting

control variables

Antibody dilutions

controls

Positive control: None specified
Negative control: None specified

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