anti phospho rpa32 rpa2 ser8  (Cell Signaling Technology Inc)

Journal: Neoplasia (New York, N.Y.)

Article Title: Suppression of ADP-ribosylation reversal triggers cell vulnerability to alkylating agents

doi: 10.1016/j.neo.2024.101092

Figure Lengend Snippet:

Article Snippet: In this study, the following antibodies were used: anti-poly/mono ADPr (rabbit monoclonal), Cell Signaling, Cat. 83732; anti-H2AX (rabbit polyclonal), Cell Signaling, Cat. 2595; anti-PARP1 (rabbit monoclonal), Abcam, Cat. ab32138; anti-PARP1 (mouse monoclonal); BD Biosciences, Cat. 556494; anti-γH2AX (mouse monoclonal), Cell Signaling, Cat. D7T2V; anti-α-tubulin (mouse monoclonal), Sigma-Aldrich, Cat. T6074; anti-β-tubulin (rabbit polyclonal), Abcam, Cat. ab6046; anti-Phospho-RPA32/RPA2 (Ser8) (rabbit polyclonal), Cell Signaling, Cat. 54762; anti-RPA32 p-S4/8 (rabbit polyclonal), Bethyl, Cat. A300-245A; anti-RPA32 (rabbit polyclonal), Bethyl, Cat. A300-244A; RPA32/RPA2 (rabbit polyclonal) Cell Signaling, Cat. 52448; anti-PAN-ADP-RIBOSE binding reagent, Merck Millipore, Cat. MABE1016; anti-Poly(ADP-ribose) (rabbit polyclonal), Enzo Life Sciences, Cat. ALX-210-890A-0100; anti-ADPRHL2 (rabbit monoclonal), Sigma-Aldrich, Cat. HPA027104; anti-histone H3 (rabbit polyclonal), Millipore, Cat. 07–690; anti-p53 (mouse monoclonal), Santa Cruz, Cat. sc-126; anti-p53 K370ac, Cell Signaling; anti-p53 K382ac, Cell Signaling, Cat. 2525; anti-Caspase 3 (rabbit monoclonal), Cell Signaling, Cat. 14220; anti-Caspase 7 (rabbit monoclonal); Cell Signaling, Cat. 12827; anti-HPF1/C4orf27 (rabbit polyclonal), NovusBio; Cat. NBP1-93973; anti-H2A (rabbit polyclonal), Abcam, Cat. ab18255; anti-PARP2 (mouse monoclonal), Millipore, Cat. MABE18; anti-lamin A (rabbit polyclonal), Abcam, Cat. ab26300;

Techniques: Binding Assay, Recombinant, Protease Inhibitor, Transfection, Proliferation Assay, Flow Cytometry, Fractionation, Cell Culture, Plasmid Preparation, Software, Microscopy

Journal: Neoplasia (New York, N.Y.)

Article Title: Suppression of ADP-ribosylation reversal triggers cell vulnerability to alkylating agents

doi: 10.1016/j.neo.2024.101092

Figure Lengend Snippet:

Article Snippet: In this study, the following antibodies were used: anti-poly/mono ADPr (rabbit monoclonal), Cell Signaling, Cat. 83732; anti-H2AX (rabbit polyclonal), Cell Signaling, Cat. 2595; anti-PARP1 (rabbit monoclonal), Abcam, Cat. ab32138; anti-PARP1 (mouse monoclonal); BD Biosciences, Cat. 556494; anti-γH2AX (mouse monoclonal), Cell Signaling, Cat. D7T2V; anti-α-tubulin (mouse monoclonal), Sigma-Aldrich, Cat. T6074; anti-β-tubulin (rabbit polyclonal), Abcam, Cat. ab6046; anti-Phospho-RPA32/RPA2 (Ser8) (rabbit polyclonal), Cell Signaling, Cat. 54762; anti-RPA32 p-S4/8 (rabbit polyclonal), Bethyl, Cat. A300-245A; anti-RPA32 (rabbit polyclonal), Bethyl, Cat. A300-244A; RPA32/RPA2 (rabbit polyclonal) Cell Signaling, Cat. 52448; anti-PAN-ADP-RIBOSE binding reagent, Merck Millipore, Cat. MABE1016; anti-Poly(ADP-ribose) (rabbit polyclonal), Enzo Life Sciences, Cat. ALX-210-890A-0100; anti-ADPRHL2 (rabbit monoclonal), Sigma-Aldrich, Cat. HPA027104; anti-histone H3 (rabbit polyclonal), Millipore, Cat. 07–690; anti-p53 (mouse monoclonal), Santa Cruz, Cat. sc-126; anti-p53 K370ac, Cell Signaling; anti-p53 K382ac, Cell Signaling, Cat. 2525; anti-Caspase 3 (rabbit monoclonal), Cell Signaling, Cat. 14220; anti-Caspase 7 (rabbit monoclonal); Cell Signaling, Cat. 12827; anti-HPF1/C4orf27 (rabbit polyclonal), NovusBio; Cat. NBP1-93973; anti-H2A (rabbit polyclonal), Abcam, Cat. ab18255; anti-PARP2 (mouse monoclonal), Millipore, Cat. MABE18; anti-lamin A (rabbit polyclonal), Abcam, Cat. ab26300;

Techniques: Binding Assay, Recombinant, Protease Inhibitor, Transfection, Proliferation Assay, Flow Cytometry, Fractionation, Cell Culture, Plasmid Preparation, Software, Microscopy

Journal: iScience

Article Title: Bromodomain protein BRD8 regulates cell cycle progression in colorectal cancer cells through a TIP60-independent regulation of the pre-RC complex

doi: 10.1016/j.isci.2023.106563

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-Phospho-RPA32/RPA2 (Ser8) , Cell Signaling Technology , Cat# 54762; RRID:AB_2799471.

Techniques: Recombinant, Plasmid Preparation, Cell Cycle Assay, CCK-8 Assay, Flow Cytometry, Viability Assay, Expressing, Clone Assay, Mutagenesis, Software

Journal: bioRxiv

Article Title: Genomes of the Autonomous Parvovirus Minute Virus of Mice Induce Replication Stress Through RPA Exhaustion

doi: 10.1101/2023.02.13.528428

Figure Lengend Snippet: (A) Schematic of the Single-Molecule DNA Fiber Assay where RPA is overexpressed in A9 cells prior to infection. Mouse A9 fibroblasts were synchronized in Isoleucine deficient media during which they were transfected with an RPA expression vector at 12 hours post-initiation of synchronization. Cells were released into complete DMEM media at 42 hours when they were concurrently infected with UV-MVM at an MOI of 25. Cells were then infected with UV-MVM at an MOI of 25 for 24 hours. Cells were pulsed with IdU and CldU before completing the rest of the DFA protocol. (B, C) Each datapoint represents the length of a single IdU and/or CldU labelled DNA fiber. The median length of at least 150 measurements of IdU and CldU tracks are represented by red and green horizontal bars respectively. The experiment was performed as described by the schematic depicted in (A). At least 150 individual fibers were measured for each condition. Similar results were obtained for three independent biological replicates of MVM infection. Statistical significance of the dataset was determined by Mann Whitney Wilcoxon test, **** represents P ≤ 0.0001. (D) Categorization of DNA fiber types as percentages out of a total of 100% as determined by presence of IdU or CldU, divided into percentages that are progressing replication forks (black), stalled replication forks (red) and new origins (green). (E) Western blot analysis of MVM infection during ectopic expression of RPA32 showing cellular levels of NS1 as a marker for virus replication, phospho-RPA32 levels due to expression and induction of cellular DNA damage and total Tubulin levels in the cell as loading control for the immunoblot.

Article Snippet: Antibodies used for western blot analysis were: Tubulin (Millipore Sigma, clone DM1A, 05-829), NS1 (2C9b monoclonal antibody), phosphor-RPA32 (Cell Signaling, 83745), γH2AX (Abcam, ab11174), HRP-conjugated anti-mouse secondary (Cell Signaling, 7076S), HRP conjugated anti-rabbit secondary (Cell Signaling, 7074S).

Techniques: Infection, Transfection, Expressing, Plasmid Preparation, MANN-WHITNEY, Western Blot, Marker