rabbit anti human brca2  (Bethyl)

Journal: Nucleic Acids Research

Article Title: Noncanonical NF-κB factor p100/p52 regulates homologous recombination and modulates sensitivity to DNA-damaging therapy

doi: 10.1093/nar/gkac491

Figure Lengend Snippet: Targeting of p100/p52 specifically sensitizes HR-proficient cells to DNA-damaging treatments. Clonogenic survival is plotted for DLD-1 cells and an isogenic BRCA2−/− derivative incubated continuously with ( A ) indicated doses of camptothecin or treated with a ( B ) single dose of ionizing radiation. Error bars represent SEM for three replicates.

Article Snippet: The following primary antibodies were used: mouse anti-human NF-κB1 p105/p50 (Santa Cruz 8414), mouse anti-human NF-κB2 p100/p52 (Millipore 05–361), rabbit anti-human RelB (Cell Signaling Technology 4922), rabbit anti-human RAD51 (Pacific Immunology, affinity purified from serum and used at 1:1000), mouse anti-human 53BP1 (Upstate 05-726 Clone BP13), rabbit anti-human H2A.X (Abcam ab229914), mouse anti-human γH2A.X (Millipore 05–636), rabbit anti-human BRCA2 (Bethyl A303-434A), rabbit anti-human GAPDH (Cell Signaling Technology 5174), and mouse anti-chicken α-tubulin (Fitzgerald 10R-T130A).

Techniques: Incubation

Journal: Oncology Letters

Article Title: Expression characteristics of FHIT , p53 , BRCA2 and MLH1 in families with a history of oesophageal cancer in a region with a high incidence of oesophageal cancer

doi: 10.3892/ol.2014.2682

Figure Lengend Snippet: Breast cancer type 2 susceptibility protein (BRCA2) immune response in oesophageal carcinoma tissues. (A) Strong and positive reaction for BRCA2 expression. Immunoreactivity was predominantly located in the cytoplasm and cell membrane; a positive cancer cell is shown by the arrow. (B) Weak and positive reaction for BRCA2 expression. Immunoreactivity was predominantly located in the cytoplasm and membrane; a positive cancer cell is shown by the arrow. (C) Negative expression for BRCA2; a negatively expressed cell is shown by the arrow. Magnification, ×400; haematoxylin and eosin restained.

Article Snippet: Subsequently, 0.5% H 2 O 2 was added to MLH1 at room temperature for 20 min and the resulting mixture was washed three times with PBS for 5 min. BRCA2 rabbit anti-human polyclonal antibody was purchased from Boster Biological Engineering Co., Ltd., (Wuhan, China; dilution 1:100), MLH1 rat anti-horse monoclonal antibody was purchased from BD Pharmingen (San Diego, CA, USA; dilution, 1:50), the FHIT polyclonal rabbit anti-goat antibody was purchased from Beijing Zhongshan Biotechnology Co., Ltd. (Beijing, China) (ZA-0410; 1:100 dilution) and p53 rat anti-horse monoclonal antibody (1:1,000) was purchased from Nuclea Biotechnologies, Inc., (Pittsfield, MA, USA).

Techniques: Expressing, Membrane

Journal: Oncology Letters

Article Title: Expression characteristics of FHIT , p53 , BRCA2 and MLH1 in families with a history of oesophageal cancer in a region with a high incidence of oesophageal cancer

doi: 10.3892/ol.2014.2682

Figure Lengend Snippet: Correlation analysis of BRCA2 , p53 , MLH1 and FHIT expression in the cancer tissues of oesophageal carcinoma patients with or without a family history of oesophageal cancer.

Article Snippet: Subsequently, 0.5% H 2 O 2 was added to MLH1 at room temperature for 20 min and the resulting mixture was washed three times with PBS for 5 min. BRCA2 rabbit anti-human polyclonal antibody was purchased from Boster Biological Engineering Co., Ltd., (Wuhan, China; dilution 1:100), MLH1 rat anti-horse monoclonal antibody was purchased from BD Pharmingen (San Diego, CA, USA; dilution, 1:50), the FHIT polyclonal rabbit anti-goat antibody was purchased from Beijing Zhongshan Biotechnology Co., Ltd. (Beijing, China) (ZA-0410; 1:100 dilution) and p53 rat anti-horse monoclonal antibody (1:1,000) was purchased from Nuclea Biotechnologies, Inc., (Pittsfield, MA, USA).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Expression characteristics of FHIT , p53 , BRCA2 and MLH1 in families with a history of oesophageal cancer in a region with a high incidence of oesophageal cancer

doi: 10.3892/ol.2014.2682

Figure Lengend Snippet: Correlation analysis of BRCA2 , p53 and MLH1 expression in the cancer tissues of oesophageal carcinoma patients with or without a family history of oesophageal cancer.

Article Snippet: Subsequently, 0.5% H 2 O 2 was added to MLH1 at room temperature for 20 min and the resulting mixture was washed three times with PBS for 5 min. BRCA2 rabbit anti-human polyclonal antibody was purchased from Boster Biological Engineering Co., Ltd., (Wuhan, China; dilution 1:100), MLH1 rat anti-horse monoclonal antibody was purchased from BD Pharmingen (San Diego, CA, USA; dilution, 1:50), the FHIT polyclonal rabbit anti-goat antibody was purchased from Beijing Zhongshan Biotechnology Co., Ltd. (Beijing, China) (ZA-0410; 1:100 dilution) and p53 rat anti-horse monoclonal antibody (1:1,000) was purchased from Nuclea Biotechnologies, Inc., (Pittsfield, MA, USA).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Expression characteristics of FHIT , p53 , BRCA2 and MLH1 in families with a history of oesophageal cancer in a region with a high incidence of oesophageal cancer

doi: 10.3892/ol.2014.2682

Figure Lengend Snippet: Association between BRCA2, MLH1 , FHIT and p53 positive expression and clinicopathological characteristics of oesophageal carcinoma.

Article Snippet: Subsequently, 0.5% H 2 O 2 was added to MLH1 at room temperature for 20 min and the resulting mixture was washed three times with PBS for 5 min. BRCA2 rabbit anti-human polyclonal antibody was purchased from Boster Biological Engineering Co., Ltd., (Wuhan, China; dilution 1:100), MLH1 rat anti-horse monoclonal antibody was purchased from BD Pharmingen (San Diego, CA, USA; dilution, 1:50), the FHIT polyclonal rabbit anti-goat antibody was purchased from Beijing Zhongshan Biotechnology Co., Ltd. (Beijing, China) (ZA-0410; 1:100 dilution) and p53 rat anti-horse monoclonal antibody (1:1,000) was purchased from Nuclea Biotechnologies, Inc., (Pittsfield, MA, USA).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Expression characteristics of FHIT , p53 , BRCA2 and MLH1 in families with a history of oesophageal cancer in a region with a high incidence of oesophageal cancer

doi: 10.3892/ol.2014.2682

Figure Lengend Snippet: BRCA2 and MLH1 expression analysis in patients with a positive or negative family history of oesophageal carcinoma.

Article Snippet: Subsequently, 0.5% H 2 O 2 was added to MLH1 at room temperature for 20 min and the resulting mixture was washed three times with PBS for 5 min. BRCA2 rabbit anti-human polyclonal antibody was purchased from Boster Biological Engineering Co., Ltd., (Wuhan, China; dilution 1:100), MLH1 rat anti-horse monoclonal antibody was purchased from BD Pharmingen (San Diego, CA, USA; dilution, 1:50), the FHIT polyclonal rabbit anti-goat antibody was purchased from Beijing Zhongshan Biotechnology Co., Ltd. (Beijing, China) (ZA-0410; 1:100 dilution) and p53 rat anti-horse monoclonal antibody (1:1,000) was purchased from Nuclea Biotechnologies, Inc., (Pittsfield, MA, USA).

Techniques: Expressing

Journal:

Article Title: Radiation Enhances Caspase 3 Cleavage of Rad51 in BRCA2-Defective Cells

doi: 10.1667/RR1129.1

Figure Lengend Snippet: Structure and expression of BRCA2. Panel A: Human BRCA2 contains 3418 amino acids. The central region contains a series of eight conserved BRC repeats that bind RAD51. The 6174delT mutation produces a frameshift and premature termination that disrupts the last two BRC repeats and introduces a premature stop codon. Panel B: Western blot of 35S-labeled cells with anti-human C-terminal BRCA2 antibody. Lane 1: untransfected Capan-1 cells; lane 2: Capan-1 cells transfected with human BRCA2 cDNA; lane 3: HeLa cells.

Article Snippet: We used affinity-purified rabbit anti-human RAD51 (Ab-1, Oncogene Research Products) at a dilution of 1:2500, affinity-purified rabbit anti-human BRCA2 (anti-BRCA2, amino acids 3245–3418, Pharmingen) at a dilution of 1:1000, and affinity-purified rabbit anti-human caspase 3 antibody (Pharmingen) at a dilution of 1:2000 as primary antibodies.

Techniques: Expressing, Mutagenesis, Western Blot, Labeling, Transfection

Journal:

Article Title: Radiation Enhances Caspase 3 Cleavage of Rad51 in BRCA2-Defective Cells

doi: 10.1667/RR1129.1

Figure Lengend Snippet: Panel A: Caspase 3 cleavage of RAD51 after irradiation. Western blot showing caspase 3-mediated cleavage of RAD51 (large arrow, 38 kDa) to RAD51-C3 (small arrow, 21 kDa) after exposure to 10 Gy of ionizing radiation (time in hours). NI is the nonirradiated control; 0.1 represents an aliquot processed 6 min after irradiation (i.e., as rapidly as possible). Polyclonal RAD51 antibody detects the 38-kDa RAD51 and 21-kDa cleavage product. Panel 1, Capan-1 cells; panel 2, Capan-1 cells transfected with a full-length human BRCA2 cDNA; panel 3, Brca2− cells; panel 4, Brca2− cells transfected with BRCA2 cDNA; panel 5, MCF-7 cells, which reportedly lack caspase 3 activity (9). Samples were standardized by loading equal concentrations of protein. Panel B: Caspase 3 immunoprecipitation (IP)-Western blot showing association of Rad51 with caspase 3 before and after irradiation (time in hours). Immunoprecipitation with Rad51 antibody, immunoprecipitates fractionated by SDS-PAGE and blotted with a 1:2000 dilution of caspase 3 antibody. Upper panel, Capan-1 cells; lower panel, BRCA2-transfected Capan-1 cells, standardized by loading equal protein. Arrow indicates 32-kDa human caspase 3 protein. Panel C: Rad51 IP-Western blot. Immunoprecipitation with caspase 3 antibody, then blotting with Rad51 antibody (1:2500). Arrow indicates the 36-kDa human Rad51 protein. In panels B and C, the darker line denotes heavy-chain and the lighter line light-chain immunoglobulin bands.

Article Snippet: We used affinity-purified rabbit anti-human RAD51 (Ab-1, Oncogene Research Products) at a dilution of 1:2500, affinity-purified rabbit anti-human BRCA2 (anti-BRCA2, amino acids 3245–3418, Pharmingen) at a dilution of 1:1000, and affinity-purified rabbit anti-human caspase 3 antibody (Pharmingen) at a dilution of 1:2000 as primary antibodies.

Techniques: Irradiation, Western Blot, Transfection, Activity Assay, Immunoprecipitation, SDS Page

Journal:

Article Title: Radiation Enhances Caspase 3 Cleavage of Rad51 in BRCA2-Defective Cells

doi: 10.1667/RR1129.1

Figure Lengend Snippet: Caspase 3 activity in irradiated BRCA2-defective and transfected cells. panel A: Caspase 3 activity in irradiated Capan-1 cells by identical assay. Panel B: Caspase 3 activity in irradiated Brca2− cells. Panel C: Caspase 3 activity in irradiated mouse embryo fibroblasts. OD on the Y axis is caspase 3 activity as measured by enzymatic conversion of DEVD-p-nitroaniline to p-nitroaniline measured at 405 nm. Points are triplicate determinations.

Article Snippet: We used affinity-purified rabbit anti-human RAD51 (Ab-1, Oncogene Research Products) at a dilution of 1:2500, affinity-purified rabbit anti-human BRCA2 (anti-BRCA2, amino acids 3245–3418, Pharmingen) at a dilution of 1:1000, and affinity-purified rabbit anti-human caspase 3 antibody (Pharmingen) at a dilution of 1:2000 as primary antibodies.

Techniques: Activity Assay, Irradiation, Transfection