rabbit polyclonal  (Novus Biologicals)

Journal: PLoS ONE

Article Title: The Role of Nibrin in Doxorubicin-Induced Apoptosis and Cell Senescence in Nijmegen Breakage Syndrome Patients Lymphocytes

doi: 10.1371/journal.pone.0104964

Figure Lengend Snippet: A. Level of nibrin: wild-type (p95) and the truncated form (p70) in control (C) and doxorubicin treated (D, 1 µM/1 h) S3R, S4 and VSMCs. Expression of nibrin was analyzed by immunoprecipitation using an anti-NBS1 antibody followed by Western blotting with anti-NBS1 (upper panel). Alternatively, IP using anti-MRE11 antibody was performed followed by WB with anti-NBS1 (lower panel). MRE11 was used as a loading control. The last lane (C IP) shows the negative IP control. Note that p95 is only present in VSMCs, in which there is no p70-nibrin. B. ATM binding to nibrin in control and doxorubicin-treated S3R and S4 cells analyzed by immunoprecipitation using anti-NBS1 antibody (upper panel) or anti-ATM antibody (lower panel). Levels of ATM and p70 were detected by WB. Loading controls were performed in both variants of IP. C. Expression of BRCA1 in control and dox-treated S3R and S4 cells was analyzed by immunoprecipitation using anti-ATM antibody followed by WB using an anti-BRCA1 antibody. Loading and negative IP controls were performed as above.

Article Snippet: Mre11, BRCA1, ATM and NBS1 were detected using the Western blotting technique with the following antibodies: BRCA1 mouse monoclonal (R&D, Biokom, Warsaw, Poland), ATM rabbit monoclonal antibody (Epitomics, Burlingame, California, USA), Mre11 rabbit monoclonal antibody (Cell Signaling, Lab-JOT Ltd., Warsaw, Poland), NBS1 rabbit polyclonal antibody (Sigma Aldrich, Poznan, Poland) and secondary rabbit polyclonal antibody conjugated with horseradish peroxidase (Dako, Poland).

Techniques: Expressing, Immunoprecipitation, Western Blot, Binding Assay

Journal: PLoS ONE

Article Title: The Role of Nibrin in Doxorubicin-Induced Apoptosis and Cell Senescence in Nijmegen Breakage Syndrome Patients Lymphocytes

doi: 10.1371/journal.pone.0104964

Figure Lengend Snippet: Cells were transfected with negative siRNA (−) or NBN siRNA (+) and afterwards cultured for three days in the presence of doxorubicin (100 nM). A. Downregulation of the NBS1 protein level in VSMCs using specific siRNA (60 nM). Whole cell extracts were prepared at indicated time points after treatment with doxorubicin. Expression of the indicated proteins was estimated by Western blotting, β-actin was used as a loading control. The amount of the protein in cells transfected with NBN siRNA was calculated by densitometry as a fraction of that present in cells transfected with negative siRNA (1). B. 53BP1 staining in doxorubicin-treated control cells and cells with silenced nibrin. Representative images from one of three independent experiments. Magnification 100x. C. 53BP1 staining in doxorubicin treated control cells and cells with silenced nibrin. Cells with DNA damage were divided into four groups based on the number of 53BP1 foci: cells without 53BP1 foci, with one focus, with 2–5 foci, with more than 5 foci. Means from three independent experiments. D. SA-β-Gal activity in doxorubicin treated VSMC cells. Representative images from one of three independent experiments, magnification 100x. E. The percentage of SA-β-Gal positive cells (a mean ± SD) from three independent experiments. F. BrdU incorporation assay. Control cells and cells transfected with negative siRNA or NBN si RNA were cultured with BrdU for 24 h. Data presented as means ± SD from three independent experiments.

Article Snippet: Mre11, BRCA1, ATM and NBS1 were detected using the Western blotting technique with the following antibodies: BRCA1 mouse monoclonal (R&D, Biokom, Warsaw, Poland), ATM rabbit monoclonal antibody (Epitomics, Burlingame, California, USA), Mre11 rabbit monoclonal antibody (Cell Signaling, Lab-JOT Ltd., Warsaw, Poland), NBS1 rabbit polyclonal antibody (Sigma Aldrich, Poznan, Poland) and secondary rabbit polyclonal antibody conjugated with horseradish peroxidase (Dako, Poland).

Techniques: Transfection, Cell Culture, Expressing, Western Blot, Staining, Activity Assay, BrdU Incorporation Assay

Journal: Cell Reports

Article Title: Protein phosphatase 1 acts as a RIF1 effector to suppress DSB resection prior to Shieldin action

doi: 10.1016/j.celrep.2021.109383

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-NBS1 , Novus Biologicals , Cat# NB 100-143 RRID: AB_350080.

Techniques: In Situ, Proximity Ligation Assay, Recombinant, Electron Microscopy, Imaging, Knock-Out, Control, Plasmid Preparation, Expressing, Software