For whole-cell extract preparation, cells were lysed in SDS lysis buffer (50 mM Tris pH7.5, 20 mM NaCl, 10 mM MgCl2, 0.1% SDS, anti-proteases) supplemented with benzonase for 10 min at room temperature, as previously described32 (link). Proteins were quantified with Bradford assay. Proteins were separated on 8 or 15% acrylamide SDS–polyacrylamide gel electrophoresis. Membranes were blotted with antibodies directed the following proteins: β-actin (mouse AC-15, Sigma #A5441, 1/10,000), Flag (mouse M2, Sigma #F4049, 1/1,000), GFP (mouse, Roche #11814460001, 1/1,000), histone H2B (rabbit V119, Cell Signaling #8135, 1/1,000), histone H4 (mouse, Abcam #ab31830, 1/1,000), HA (mouse HA.11 16B12, Covance #MMS-101R, 1/1,000), 6x-His tag (mouse #631212, Clontech, 1/5,000), PCNA (mouse PC10, Santa Cruz #sc-56, 1/4,000), Ub-PCNA Lys164 (rabbit D5C7P, Cell Signaling #13439, 1/1,000), PIAS1 (rabbit, Epitomics #2474, 1/5,000), PIAS4 (rabbit, ProteinTech #14242-1-AP, 1/1,000), polδ-p125 (goat C-20, Santa Cruz #sc-8797, 1/1,000), polη (rabbit, Abcam #ab17725, 1/1,000; rabbit H-300, Santa Cruz #sc-5592, 1/2,000; mouse B-7, Santa Cruz #sc-17770, 1/500; rabbit, Bethyl #A301-231A, 1/1,000), Rad18 (mouse, Abcam #ab57447; rabbit, Bethyl #A301-340A, 1/2,000), RPA32 (mouse, Calbiochem #NA19L, 1/5,000). Uncropped images for the most relevant blots are shown in Supplementary Fig. 11 .
Ab57447
Manufactured by Abcam
Ab57447 is a laboratory product offered by Abcam. It is a tool used for research purposes. The core function of this product is to serve as a reagent or component in scientific experiments and investigations. No further details about the intended use or specific applications of this product are provided.
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Lab products found in correlation
Ab17725, by Santa Cruz Biotechnology (1 mentions)
Nb100 182, by Novus Biologicals (1 mentions)
H 300, by Santa Cruz Biotechnology (1 mentions)
M1445, by Merck Group (1 mentions)
Ab92312, by Abcam (1 mentions)
Ab1790, by Abcam (1 mentions)
Ab17725, by Abcam (1 mentions)
Ab290, by Abcam (1 mentions)
D5c7p, by Cell Signaling Technology (1 mentions)
Ab16048, by Abcam (1 mentions)
2 protocols using ab57447
1
Whole-Cell Extract Preparation and Western Blot Analysis
2
Antibody Detection Protocol for DNA Repair Factors
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Antibodies to the following proteins were used in this study: sheep α-FAN1 (1:500, a kind gift from John Rouse, University of Dundee), rabbit α-FANCD2 (1:5000, NB100-182, Novus Biologicals), mouse α-PCNA (1:1000, PC10, SantaCruz), rabbit α-ubiquitylated-PCNA (1:1000, D5C7P, Cell Signaling), mouse α-RAD18 (1:500, ab57447, Abcam), rabbit α-MLH1 (1:5000, ab92312, Abcam), rabbit α-Polη (1:1000, ab17725, Abcam), rabbit α-REV1 (1:1000, H-300, SantaCruz), rabbit α-GFP (1:1000, ab290, Abcam), rabbit α-LaminB1 (1:1000, ab16048, Abcam), rabbit α-histone H2B (1:1000, ab1790, Abcam), mouse α−MUS81 (1:1000, M1445, Sigma Aldrich), rabbit α−USP1 (1:1000, Bethyl labs. A301-700A).
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