rabbit polyclonal anti rad51 primary antibodies  (Santa Cruz Biotechnology)

Journal: Oncology Reports

Article Title: M2 macrophages reduce the radiosensitivity of head and neck cancer by releasing HB-EGF

doi: 10.3892/or.2020.7628

Figure Lengend Snippet: In CAL27 cells, inhibition of EGFR by small molecule inhibitor results in reduced NHEJ repair capacity. (A) CAL27 and CAL27 with HB-EGF (50 ng/ml) were assessed by the western blot analysis with anti-EGFR and anti-p-EGFR antibodies. (B) EGFR and EGFR phosphorylation expression in CAL27 cells with or without HB-EGF after 4 Gy radiation. NC, CAL27 cells. (C) Representative images of the number of the p-DNA-PK foci per cell in 5 h after irradiation (4 Gy) in CAL27 cells with or without the EGFR inhibitor; magnification, ×200 (green, p-DNA-PK staining; blue, DAPI). (D) Representative images of the number of the RAD51 foci per cell in 5 h after irradiation (4 Gy) in CAL27 cells with or without the EGFR inhibitor; magnification ×200 (green, p-DNA-PK staining; blue, DAPI). *P<0.05. In CAL27 cells, inhibition of EGFR by small molecule inhibitor results in reduced NHEJ repair capacity. (E-a) Number of p-DNA-PK foci of NHEJ specific markers in CAL27 cells with or without EGFR inhibitors; (E-b) Number of RAD51 foci of HR specific markers in CAL27 cells with or without EGFR inhibitors. (F) IHC analyses of EGFR in cancer and normal tissues; magnification, ×400 (HPV + T, HPV-positive tumor tissues; HPV-T, HPV-negative tumor tissues; HPV + /HPV − N, head and neck normal tissues). (G) EGFR IHC score. HPV-positive cancer tissues (n=19), HPV-negative cases (n=33). (H) Expression of EGFR between HPV-positive (n=98) and HPV-negative (n=420) HNSCC in the TCGA database. Results are presented as the mean ± SD. *P<0.05. HPV, human papilloma virus; HNSCC, head and neck squamous cell carcinoma; EGFR, epidermal growth factor receptor; NHEJ, non-homologous end-joining; HB-EGF, heparin-binding epidermal growth factor; p-, phosphorylated; DNA-PK, DNA-dependent protein kinase; RAD51, RAD51 recombinase; IHC, immunohistochemistry; TCGA, The Cancer Genome Atlas.

Article Snippet: Samples were incubated with the γ H2A histone family member X (H2AX) mouse monoclonal antibody (product code ab26350; Abcam) at 1:500 dilution, primary rabbit polyclonal antibodies against RAD51 recombinase (RAD51; product code ab133534; Abcam) at 1:200, and phosphorylated (p)-DNA-dependent protein kinase (DNA-PK; phosphor S2056; product code ab124918; Abcam) at 1:200 and EGFR inhibitor gefitinib (ZD1839; cat. no. HY-50895; MedChemExpress) overnight at 4°C.

Techniques: Inhibition, Western Blot, Phospho-proteomics, Expressing, Irradiation, Staining, Virus, Non-Homologous End Joining, Binding Assay, Immunohistochemistry

Journal: International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group

Article Title: BRCA2 protects mammalian cells from heat shock.

doi: 10.1080/02656736.2017.1370558

Figure Lengend Snippet: Figure 1. RAD51 foci formation after heat shock. (A) Typical photographs of RAD51 in V79 cells at indicated time points after heat shock (44 C for 60 min). a–d, RAD51; e–h, DAPI; i–l, merge. (B) RAD51 foci formation at indicated times after heat shock. Columns show the mean of three independent experiments. Bars indicate the SD. (C) Immunofluorescence staining for cH2AX and RAD51 foci formation in nuclei by DAPI of V79 cells at 4 h after heat shock (44 C for 60 min) and irradiation (X-rays, 10 Gy).

Article Snippet: Then the cells were permeabilised for 5min at 4 C in 0.2% Triton X-100, and blocked in PBS with 1% Bovine Serum Albumin (BSA) for 1 h at 37 C. Cells were then incubated with anti-phospho-H2AX (Ser 139) mouse monoclonal antibodies (Upstate Biotechnology, Lake Placid, NY) at a 1:300 dilution, or rabbit polyclonal anti-RAD51 primary antibodies (H-92, Santa Cruz) for 1 h at room temperature at a 1:300 dilution in PBS containing 1% BSA.

Techniques: Immunofluorescence, Staining, Irradiation