mouse anti fancd2 antibody  (Novus Biologicals)

Journal: bioRxiv

Article Title: Aurora kinase A is a synthetic lethal target in FANCA-deficient cancers

doi: 10.64898/2026.02.04.703705

Figure Lengend Snippet: (A) Western blot illustrating FANCA protein loss in FANCA-deficient cell lines. (B,C) Lack of FANCD2 monoubiquitination, a hallmark of FA pathway activation, in FANCA-deficient cells. (D,E) FANCA-deficient cells show increased vulnerability to MMC: drug response curves for CCH-SCC-FA1 ( FANCA -/- ) vs CCH-SCC-FA1 ( FANCA Compl ) (D) and DU145 WT vs DU145_ FANCA KO (E) are illustrated. (F,G) FANCA-deficient cells show increased vulnerability to cisplatin: drug response curves for CCH-SCC-FA1 ( FANCA -/- ) vs CCH-SCC-FA1 ( FANCA Compl ) (F) and DU145 WT vs DU145_ FANCA KO (G) are illustrated.

Article Snippet: Cells were then incubated with the primary antibody for 1 hour at RT, rabbit anti-FANCD2 antibody (Novus Biologicals, NB100-182, 1:500 in SB) or mouse anti-FANCD2 antibody (Novus Biologicals, NB100-316, 1:500 in SB).

Techniques: Western Blot, Activation Assay

Journal: bioRxiv

Article Title: Aurora kinase A is a synthetic lethal target in FANCA-deficient cancers

doi: 10.64898/2026.02.04.703705

Figure Lengend Snippet: Cells were exposed to 1 µM MMC for 24 hours vs control conditions (DMSO) and imaged via confocal microscopy using Zeiss LSM 710 or Zeiss LSM 980 (63x). (A,C,E) Representative immunofluorescence images for CCH-SCC-FA1 ( FANCA Compl ) vs CCH-SCC-FA1 ( FANCA -/- ) (A), DU145 WT vs DU145_ FANCA KO (C), and RPE1 WT vs RPE1_ FANCA KD (E); scale represents 20 µm. (B, D, F) FANCD2 foci quantification for the same FANCA-deficient vs proficient cell lines. The number of foci per cell are represented, and statistical significance was calculated with the unpaired t-test.

Article Snippet: Cells were then incubated with the primary antibody for 1 hour at RT, rabbit anti-FANCD2 antibody (Novus Biologicals, NB100-182, 1:500 in SB) or mouse anti-FANCD2 antibody (Novus Biologicals, NB100-316, 1:500 in SB).

Techniques: Control, Confocal Microscopy, Immunofluorescence

Journal: Molecular Oncology

Article Title: Improving PARP inhibitor efficacy in bladder cancer without genetic BRCAness by combination with PLX51107

doi: 10.1002/1878-0261.70148

Figure Lengend Snippet: Validation of factors underlying synergism of combination treatment. (A) qRT‐PCR validation of genes after indicated treatment conditions with PLX51107 (PLX) and Olaparib (Ola) or Talazoparib (Tala) in indicated cell line pairs. TBP was measured as a housekeeping gene and used for normalization. Fold change expression (log 2 ) was calculated vs. DMSO control (set to 0). Significance is denoted in comparison to DMSO *** P ≤ 0.001, ** P ≤ 0.01, * P ≤ 0.05 (mean ± SEM; n = 3; two‐way ANOVA, Tukey's test). (B) Treatment‐induced changes of BCL2, BIRC5, and FANCD2 were further validated on the protein level by western blotting. Band intensities were quantified densitometrically using Image Lab Software (Bio‐Rad) and normalized to α‐Tubulin as loading control and DMSO as solvent control. Fold change expression (log 2 ) was calculated vs. DMSO control (set to 0). Results of three independent experiments were averaged. Significance is denoted in comparison to DMSO, *** P ≤ 0.001, ** P ≤ 0.01, * P ≤ 0.05 (mean ± SEM; n = 3; two‐way ANOVA, Tukey's test). Example western blot images are displayed in Fig. .

Article Snippet: The antibodies used for target detection were PARP1 (Invitrogen #436400; 1:1000), cleaved PARP (Cell Signaling, Danvers, MA, USA #9541; 1:500), SLFN11 (Cell Signaling #34858), OAS1 (Cell Signaling #14498), BIRC5 (Cell Signaling # 2808), BCL2 (Cell Signaling #15071), FANCD2 (Cell Signaling #16323), and α‐Tubulin (Sigma‐Aldrich, Steinheim, Germany #B512; 1:50000).

Techniques: Biomarker Discovery, Quantitative RT-PCR, Expressing, Control, Comparison, Western Blot, Software, Solvent

Journal: bioRxiv

Article Title: ATM promotes reversed fork processing during DNA interstrand cross-link repair

doi: 10.1101/2025.10.08.680157

Figure Lengend Snippet: a , Model of replication-coupled ICL repair by the FA pathway. b , Schematic describing CRISPR dropout screens using human FANCD2 +/+ or FANCD2 -/- RPE-1 cells. c , Biplot of normZ scores relating viability in psoralen and UVA treated FANCD2 +/+ (y-axis) and FANCD2 -/- (x-axis) RPE-1 cells. d , Immunoblot demonstrating efficiency of PTPA depletion in FANCD2 +/+ and FANCD2 -/- RPE-1 CRISPR knockout cell pools. Cells were transduced with lentivirus carrying non-targeting (NT) and PTPA -targeting sgRNAs. e , Relative RPE-1 cell proliferation after cisplatin treatment was measured using CellTiter-Glo reagent. FANCD2 +/+ and FANCD2 -/- RPE-1 cells were transduced with sgRNAs as in d , and cell proliferation was measured after 5 days. Data represent the mean ± standard deviation from three technical replicates.

Article Snippet: Commercial primary antibody identities and dilutions used for blotting cell samples were as follows: rabbit polyclonal anti-PTPA (human), Cell Signaling Technology #3330, 1:1,000; mouse monoclonal anti-FANCD2 (human), Santa Cruz Biotechnology #sc-20022, 1:1,000; and mouse monoclonal anti-RPA (human), Millipore Sigma #MABE285, 1:1,000.

Techniques: CRISPR, Western Blot, Knock-Out, Transduction, Standard Deviation

Journal: bioRxiv

Article Title: ATM promotes reversed fork processing during DNA interstrand cross-link repair

doi: 10.1101/2025.10.08.680157

Figure Lengend Snippet: a , Lysates from RPE-1 cells used for the chemogenomic screens described in were blotted for FANCD2 and RPA32 (loading control). b , Relative FANCD2 +/+ and FANCD2 -/- RPE-1 cell proliferation after psoralen + UVA treatment was measured using CellTiter-Glo reagent. Cell proliferation was measured after 5 days. Data represent the mean ± standard deviation from two independent experiments.

Article Snippet: Commercial primary antibody identities and dilutions used for blotting cell samples were as follows: rabbit polyclonal anti-PTPA (human), Cell Signaling Technology #3330, 1:1,000; mouse monoclonal anti-FANCD2 (human), Santa Cruz Biotechnology #sc-20022, 1:1,000; and mouse monoclonal anti-RPA (human), Millipore Sigma #MABE285, 1:1,000.

Techniques: Control, Standard Deviation

Journal: bioRxiv

Article Title: ATM promotes reversed fork processing during DNA interstrand cross-link repair

doi: 10.1101/2025.10.08.680157

Figure Lengend Snippet: a , Plasmids were replicated in egg extract supplemented with okadaic acid and the p97 inhibitor NMS-873, as indicated. Replication reactions were blotted for phospho-ATM (S1981) and H3 as in . b , pICL Pt was replicated in mock- or FANCD2-depleted egg extract supplemented with KU-55933, as indicated. Replication reactions were blotted for phospho-ATM (S1981) and H3 as in . c , pICL Pt was replicated in egg extract supplemented with [α- 32 P]dCTP and okadaic acid and the p97 inhibitor NMS-873, as indicated. Replication intermediates were analyzed as in .

Article Snippet: Commercial primary antibody identities and dilutions used for blotting cell samples were as follows: rabbit polyclonal anti-PTPA (human), Cell Signaling Technology #3330, 1:1,000; mouse monoclonal anti-FANCD2 (human), Santa Cruz Biotechnology #sc-20022, 1:1,000; and mouse monoclonal anti-RPA (human), Millipore Sigma #MABE285, 1:1,000.

Techniques:

Journal: bioRxiv

Article Title: ATM promotes reversed fork processing during DNA interstrand cross-link repair

doi: 10.1101/2025.10.08.680157

Figure Lengend Snippet: a , The replication reactions described in were supplemented with [α- 32 P]dCTP, and replication intermediates were resolved on a native agarose gel as in . b , FANCD2 immunodepletion. Egg extracts used in were resolved by SDS-PAGE and blotted for FANCD2. c , The replication reactions described in were supplemented with [α- 32 P]dCTP, and replication intermediates were resolved on a native agarose gel as in . d , pICL Pt was replicated in egg extract supplemented with NMS-873 (to prevent CMG unloading), okadaic acid, and increasing concentrations of linearized pCtrl, as indicated. Replication reactions were blotted for phospho-ATM (S1981) and H3 as in . Addition of linear DNA induces robust ATM activation. e , The pICL Pt replication reactions described in d were additionally supplemented with [α- 32 P]dCTP, and replication intermediates were resolved on a native agarose gel as in . Despite extensive ATM activation, slow figure 8 converged replication fork structures are largely refractory to resection and fork collapse. Red arrowsheads indicate linear plasmids and end-joining products that become labeled due to unscheduled DNA synthesis in extract.

Article Snippet: Commercial primary antibody identities and dilutions used for blotting cell samples were as follows: rabbit polyclonal anti-PTPA (human), Cell Signaling Technology #3330, 1:1,000; mouse monoclonal anti-FANCD2 (human), Santa Cruz Biotechnology #sc-20022, 1:1,000; and mouse monoclonal anti-RPA (human), Millipore Sigma #MABE285, 1:1,000.

Techniques: Agarose Gel Electrophoresis, Immunodepletion, SDS Page, Activation Assay, Labeling, DNA Synthesis

Journal: Cell reports

Article Title: PP2A licenses the FANCD2/FANCI complex for chromosome loading.

doi: 10.1016/j.celrep.2024.114971

Figure Lengend Snippet: Figure 1. FANCD2 is dephosphorylated by a phosphatase belonging to the PP2A family in response to ICLs (A) Schematic representation of the hypothesis that FANCD2 needs to be dephosphorylated by an unknown phosphatase for it to be relieved from the inhibitory phosphorylation mediated by CK2. Phosphorylation by CK2 reduces DNA affinity of the FANCD2/FANCI complex, preventing binding to negatively charged DNA via electrostatic repulsion. Dephosphorylation by a phosphatase is expected to alleviate the inhibition and permit chromatin loading. Parts of this figure were made in Biorender. (B) Immunoblot analysis of HeLa cells before and after treatment with 200 nM okadaic acid (OA), collected 1, 2, or 3 h post TMP/UVA treatment. (C) Live-cell imaging of HeLa FANCD2/ cells complemented with EGFP-FANCD2 and mCherry-UHRF1 treated with 20 mg/mL TMP and irradiated by a localized laser at indicated areas (white arrows), in the presence or absence of 200 nM OA. Cells were monitored for the indicated time post-irradiation. Mean ± SEM; number of cells analyzed: 5 cells for control, 5 for OA; n = 3 biological replicates. Scale bar: 10 mm.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-FANCD2 (FI17) Santa Cruz Biotechnology Cat# sc-20022; RRID: AB2278211 Rabbit polyclonal anti-FANCI FARF N/A Mouse monoclonal anti-a-Tubulin (DM1A) Sigma-Aldrich Cat# 05–829; RRID: AB310035 Mouse monoclonal anti-UHRF1 (H-8) Santa Cruz Biotechnology Cat# sc-373750; RRID: AB10947236 Mouse monoclonal anti-PPP2CA BD Biosciences Cat# 610555; RRID: AB397909) Rabbit polyclonal anti-PPP2R3A Sigma-Aldrich Cat# HPA035829 RRID: AB10696513) Rabbit multi-monoclonal anti-phospho-CK2 substrate motif [(pS/pT)DXE] Cell Signaling Cat# 8738; RRID: AB2797653 Mouse monoclonal anti-PP2A-Aa/b (PPP2R1A/PPP2R1B) (4G7) Santa Cruz Biotechnology Cat# sc13600; RRID: AB628178 Rabbit multi-monoclonal anti-phosphoATM/ATR substrate motif [(pS/pT)QG] Cell Signaling Cat# 6966; RRID: AB10949894 Rabbit anti-mouse immunoglobulins (horseradish peroxidase conjugated) Dako-Agilent Cat# P0260; RRID: AB2636929 Goat anti-rabbit immunoglobulins (horseradish peroxidase conjugated) Dako-Agilent Cat# P0448; RRID: AB2617138 Bacterial and virus strains Top10 Thermo Fisher Cat# 404010 Chemicals, peptides, and recombinant proteins FuGENE6 Promega Cat# E2691 Lipofectamine RNAiMAX Thermo Fisher Cat# 12333563 Cellfectin II Thermo Fisher Cat# 10458833 Geneticin (G418 sulfate) Thermo Fisher Cat# 10131-035 Gibco Grace’s Insect Medium, unsupplemented Thermo Fisher Cat# 11514546 Sf-900 II SFM Thermo Fisher Cat# 10902088 JF549 Promega Cat# GA1110 Protein A Sepharose Cl-4B Thermo Fisher Cat# GE17-0963-02 Dynabeads Goat Anti-Mouse IgG Thermo Fisher Cat# 11033 Anti-FLAG M2 agarose resin Sigma-Aldrich Cat# A2220 Ni2–NTA agarose resin QIAGEN Cat# 30210 Trioxsalen (TMP) Sigma-Aldrich Cat# 6137 Mitomycin C from Streptomyces caespitosus Sigma-Aldrich Cat# M4287 Benzonase Sigma-Aldrich Cat# E1014 CK2 NEB Cat# P6010S Lambda protein phosphatase NEB Cat# P0753S CX-4549 Enzo Life Sciences Cat# ENZ-CHM151-0001 Propidium iodide Sigma-Aldrich Cat# P4170 Ribonuclease A Sigma-Aldrich Cat# R5503 Critical Commercial Assays DNA sequencing Source BioScience N/A Experimental models: Cell lines HeLa FANCD2 / Lopez-Martinez et al. 10 N/A HeLa FANCD2 / + EGFP-FANCD2 + mCherry-UHRF1 Lopez-Martinez et al.10 N/A HeLa FANCD2 / + EGFP-FANCD2-6A + mCherry-UHRF1 Lopez-Martinez et al.10 N/A (Continued on next page) Cell Reports 43, 114971, November 26, 2024 17

Techniques: Phospho-proteomics, Binding Assay, De-Phosphorylation Assay, Inhibition, Western Blot, Live Cell Imaging, Irradiation, Control