anti-brca1 mouse monoclonal antibody  (Millipore)

Journal: Cell Reports Medicine

Article Title: Homologous recombination repair status in metastatic prostate cancer by next-generation sequencing and functional immunofluorescence

doi: 10.1016/j.xcrm.2025.101937

Figure Lengend Snippet: Correlation between RAD51-IF score and HRR gene alterations

Article Snippet: The following primary antibodies were used: rabbit anti-RAD51 (Abcam ab133534, 1:1,000), mouse anti-GMN (NovoCastra NCL-L, 1:60), rabbit anti-GMN (ProteinTech 10802–1-AP, 1:400), and mouse anti-γH2AX (Millipore #05636, 1:200); additionally, BRCA1 expression was assessed in selected cases using a mouse anti-BRCA1 primary antibody (Santa Cruz Biotechnology Inc, Dallas, TX sc-6954, 1:50).

Techniques:

Journal: Cell Reports Medicine

Article Title: Homologous recombination repair status in metastatic prostate cancer by next-generation sequencing and functional immunofluorescence

doi: 10.1016/j.xcrm.2025.101937

Figure Lengend Snippet: Representative cases correlating RAD51-IF score and genomic characterization Images of the RAD51 and yH2AX staining from (A) a RAD51 high sample with no HRR alterations, (B) a RAD51 low case with a BRCA2 pathogenic mutation, (C) a liver biopsy of a patient with a BRCA2 pathogenic mutation after progression to platinum-based chemotherapy that shows high percentage of cells positive for RAD51 foci; contemporaneous ctDNA analysis demonstrated BRCA2 reversion mutations. CT scan (left) of the liver lesions of the patient from baseline, response, and progression to carboplatin. Liver lesions are highlighted in yellow. Representative image of the RAD51 positivity (right) by IF and (D) prostate and liver biopsies of a patient with a monoallelic somatic BRCA1 mutation detected by NGS. The primary prostate tumor shows RAD51-negative cells, but the liver metastasis shows high RAD51 score, in parallel to BRCA1 expression by IF in this liver lesion, but not in the prostate tumor, suggesting restoration of BRCA1 expression in the metastases. Scale bar: 50 μm.

Article Snippet: The following primary antibodies were used: rabbit anti-RAD51 (Abcam ab133534, 1:1,000), mouse anti-GMN (NovoCastra NCL-L, 1:60), rabbit anti-GMN (ProteinTech 10802–1-AP, 1:400), and mouse anti-γH2AX (Millipore #05636, 1:200); additionally, BRCA1 expression was assessed in selected cases using a mouse anti-BRCA1 primary antibody (Santa Cruz Biotechnology Inc, Dallas, TX sc-6954, 1:50).

Techniques: Staining, Mutagenesis, Computed Tomography, Expressing

Journal: Cell Reports Medicine

Article Title: Homologous recombination repair status in metastatic prostate cancer by next-generation sequencing and functional immunofluorescence

doi: 10.1016/j.xcrm.2025.101937

Figure Lengend Snippet:

Article Snippet: The following primary antibodies were used: rabbit anti-RAD51 (Abcam ab133534, 1:1,000), mouse anti-GMN (NovoCastra NCL-L, 1:60), rabbit anti-GMN (ProteinTech 10802–1-AP, 1:400), and mouse anti-γH2AX (Millipore #05636, 1:200); additionally, BRCA1 expression was assessed in selected cases using a mouse anti-BRCA1 primary antibody (Santa Cruz Biotechnology Inc, Dallas, TX sc-6954, 1:50).

Techniques: Recombinant, Software

Journal: eLife

Article Title: Synergistic effect of inhibiting CHK2 and DNA replication on cancer cell growth

doi: 10.7554/eLife.104718

Figure Lengend Snippet: ( A ) MCF-7 cells treated with DMSO, 15 μM IBC, 20 μM bakuchiol (BKC), the combination IBC + BKC or AKT inhibitor, MK-2206, (AKTi, 10 μM) for 24 hr. Autophosphorylation of AKT was detected by immunoblotting analysis. Densitometric quantification of phosphor-AKT signal is shown. (n = 2). ( B ) MCF-7 cells treated overnight with DMSO, IBC, or AKT inhibitor (AKTi). Total RNA was isolated. Reverse transcription and quantitative PCR was performed with specific primers targeting the gene bodies of PCNA, E2F1, and E2F2 (n = 2). ( C ) MCF-7 cells were treated with IBC or AKT inhibitor (AKTi) for 24 hr. They were then sequentially labeled with IdU and CldU for 20 min. Replication fork progression was measured using DNA fiber spreading. The median length of CldU tracks of two biological replicates is indicated in red. ( D ) In vitro kinase assay of 43 cell cycle-related kinases following treatment with 30 µM IBC. The CHK2(I157T) mutation is linked to an increased risk of breast and colorectal cancers. CHK2(R145W) is associated with Li–Fraumeni syndrome. Both mutations do not affect the basal kinase activity of CHK2. ( E ) Percentage inhibition of kinase activity by IBC treatment is shown. ( F ) AKT kinase peptides were incubated with a half-log range dilution series of Isobavachalcone and in vitro kinase activity was measured using a radiometric assay. Data are presented as mean ± SD with a technical triplicate. ( G ) Densitometric quantification of the pCHK2-S516 induction by CPT for . The relative induction of pCHK2-S516 by CPT after IBC or BML-277 treatment compared to DMSO control is indicated. Mean ± SD of three independent experiments for DMSO + CPT and IBC + CPT are shown. The p-value was determined using two-tailed unpaired t -test. ( H ) Densitometric quantification of the pCHK2-S516 induction by CPT for . The relative induction of pBRCA1-S988 by CPT after IBC or BML-277 treatment compared to DMSO control is indicated. Mean ± SD of three independent experiments for DMSO + CPT and IBC + CPT are shown. The p-value was determined using two-tailed unpaired t -test. ( I ) Densitometric quantification of the pCHK1-S296 induction by HU for . The relative induction of pCHK1-S296 by HU after IBC or BML-277 treatment compared to DMSO control is indicated (n = 2). Figure 4—figure supplement 1—source data 1. Original membranes corresponding to . 4 and 7 are our codes for BKC and IBC, respectively. d: DMSO control; 7+4: means BKC + IBC. Figure 4—figure supplement 1—source data 2. Original membranes corresponding to with labels.

Article Snippet: Antibody , Mouse monoclonal anti-pBRCA1 (S988) , sc-166793 , Santa Cruz , 1/200.

Techniques: Western Blot, Isolation, Reverse Transcription, Real-time Polymerase Chain Reaction, Labeling, In Vitro, Kinase Assay, Mutagenesis, Activity Assay, Inhibition, Incubation, Control, Two Tailed Test

Journal: eLife

Article Title: Synergistic effect of inhibiting CHK2 and DNA replication on cancer cell growth

doi: 10.7554/eLife.104718

Figure Lengend Snippet: ( A ) Selected protein kinases were incubated with the indicated range of IBC concentrations, and kinase activity in vitro was determined by using a radiometric assay. The IC 50 of IBC for each kinase is indicated in the panel on the right. ( B ) MCF-7 cells were pretreated with DMSO, 15 µM IBC, or 20 µM BML-277 for 2 hr, then camptothecin (CPT, 1 μM) was added for 2 hr. Phosphorylation of CHK2 on S516 (pCHK2) was detected by western blotting. The ratio of pCHK2-S516 induction, relative to the DMSO + CPT control, is indicated (n = 3). ( C ) MCF-7 cells were treated as indicated in (B). Phosphorylation of chromatin-bound BRCA1 at residue S988 was detected by western blotting. The relative ratio of pBRCA1-S988 signal, after normalization to Ponceau signal, is indicated. TBP was used as a marker of chromatin fraction. ( D ) MCF-7 cells were treated with 15 μM IBC for 2 hr, then 4 mM HU was added for 2 hr. CHK1 autophosphorylation on S296 (pCHK1) was detected by western blotting. ( E, F ) In silico molecular docking of IBC in the active sites of CHK2 and CHK1, respectively. ( G, H ) Cellular thermal shift assay (CETSA) of IBC on the thermal stability of CHK2 and CHK1. MCF-7 cells were treated with 15 µM IBC or 20 μΜBML-277 for 2 hr. Cells were proceeded to CETSA as described in the ‘Materials and methods’. The amount of CHK2 and CHK1 present in the supernatant was detected by western blotting. The relative CHK2 and CHK1 signal was quantified. The p-values were determined using two-tailed paired t -test (n = 3). Figure 4—source data 1. Original membranes corresponding to with labels. Figure 4—source data 2. Original membranes corresponding to .

Article Snippet: Antibody , Mouse monoclonal anti-pBRCA1 (S988) , sc-166793 , Santa Cruz , 1/200.

Techniques: Incubation, Activity Assay, In Vitro, Phospho-proteomics, Western Blot, Control, Residue, Marker, In Silico, Thermal Shift Assay, Two Tailed Test

Journal: eLife

Article Title: Synergistic effect of inhibiting CHK2 and DNA replication on cancer cell growth

doi: 10.7554/eLife.104718

Figure Lengend Snippet:

Article Snippet: Antibody , Mouse monoclonal anti-pBRCA1 (S988) , sc-166793 , Santa Cruz , 1/200.

Techniques: