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mouse-α-brca2 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc mouse-α-brca2 antibody
    ( A ) A549 tumor cells treated with <t>BRCA2</t> siRNA were irradiated with 1 Gy (0.5 h) or 2 Gy (8 h) and immunostained with the indicated antibodies. Using EdU and cell cycle markers to distinguish G1- from G2-phase cells , focal intensities of pATM were measured using ImageJ software (see ). BRCA2 siRNA was used in this analysis to accumulate resected DSBs. ( B ) 2BN hTert (XLF-deficient) human fibroblasts were analyzed 2 h post IR with 1 Gy. Cells were stained against γH2AX and RAD51 or pATM and RAD51, and γH2AX or pATM focal intensities were measured at RAD51-foci-positive or RAD51-foci-negative foci. XLF-deficient cells were used in this analysis to prevent repair of EC DSBs during the time needed for resection of HC DSBs. ( C ) 82-6 hTert (wt) and F02-98 hTert (ATR-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). ( D ) 2BN hTert (XLF-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). Since both DNA-PK and ATM can phosphorylate γH2AX, cells were treated with DNA-PK inhibitor under all conditions. Inhibitors were added 1 h post IR, a time sufficient to allow for ATM-dependent resection and RAD51 loading (see ). In (A–D), at least 300 foci from 3 independent experiments were analyzed for each point. Box plots were used with a maximum whisker-length of 1.5-fold the inter-quartile range; the lower and upper “x” indicates the 1% or 99% margin of the data range, respectively.
    Mouse α Brca2 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse-%CE%B1-brca2+antibody/anti+brca2/pmc03731223-231-17-20
    Average 90 stars, based on 1 article reviews
    mouse-α-brca2 antibody - by Bioz Stars, 2026-08
    90/100 stars

    Images

    1) Product Images from "ATM Release at Resected Double-Strand Breaks Provides Heterochromatin Reconstitution to Facilitate Homologous Recombination"

    Article Title: ATM Release at Resected Double-Strand Breaks Provides Heterochromatin Reconstitution to Facilitate Homologous Recombination

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1003667

    ( A ) A549 tumor cells treated with BRCA2 siRNA were irradiated with 1 Gy (0.5 h) or 2 Gy (8 h) and immunostained with the indicated antibodies. Using EdU and cell cycle markers to distinguish G1- from G2-phase cells , focal intensities of pATM were measured using ImageJ software (see ). BRCA2 siRNA was used in this analysis to accumulate resected DSBs. ( B ) 2BN hTert (XLF-deficient) human fibroblasts were analyzed 2 h post IR with 1 Gy. Cells were stained against γH2AX and RAD51 or pATM and RAD51, and γH2AX or pATM focal intensities were measured at RAD51-foci-positive or RAD51-foci-negative foci. XLF-deficient cells were used in this analysis to prevent repair of EC DSBs during the time needed for resection of HC DSBs. ( C ) 82-6 hTert (wt) and F02-98 hTert (ATR-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). ( D ) 2BN hTert (XLF-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). Since both DNA-PK and ATM can phosphorylate γH2AX, cells were treated with DNA-PK inhibitor under all conditions. Inhibitors were added 1 h post IR, a time sufficient to allow for ATM-dependent resection and RAD51 loading (see ). In (A–D), at least 300 foci from 3 independent experiments were analyzed for each point. Box plots were used with a maximum whisker-length of 1.5-fold the inter-quartile range; the lower and upper “x” indicates the 1% or 99% margin of the data range, respectively.
    Figure Legend Snippet: ( A ) A549 tumor cells treated with BRCA2 siRNA were irradiated with 1 Gy (0.5 h) or 2 Gy (8 h) and immunostained with the indicated antibodies. Using EdU and cell cycle markers to distinguish G1- from G2-phase cells , focal intensities of pATM were measured using ImageJ software (see ). BRCA2 siRNA was used in this analysis to accumulate resected DSBs. ( B ) 2BN hTert (XLF-deficient) human fibroblasts were analyzed 2 h post IR with 1 Gy. Cells were stained against γH2AX and RAD51 or pATM and RAD51, and γH2AX or pATM focal intensities were measured at RAD51-foci-positive or RAD51-foci-negative foci. XLF-deficient cells were used in this analysis to prevent repair of EC DSBs during the time needed for resection of HC DSBs. ( C ) 82-6 hTert (wt) and F02-98 hTert (ATR-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). ( D ) 2BN hTert (XLF-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). Since both DNA-PK and ATM can phosphorylate γH2AX, cells were treated with DNA-PK inhibitor under all conditions. Inhibitors were added 1 h post IR, a time sufficient to allow for ATM-dependent resection and RAD51 loading (see ). In (A–D), at least 300 foci from 3 independent experiments were analyzed for each point. Box plots were used with a maximum whisker-length of 1.5-fold the inter-quartile range; the lower and upper “x” indicates the 1% or 99% margin of the data range, respectively.

    Techniques Used: Irradiation, Software, Staining, Whisker Assay

    ( A ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) and HSC62 hTert (BRCA2-deficient) human fibroblasts. ( B ) RPA and RAD51 foci analysis in G2-irradiated A549 tumor cells. ( C ) Endogenous KAP-1 and BRCA2 was depleted in HeLa tumor cells by siRNA, and cells were transfected with GFP-tagged and siRNA-resistant empty (GFP), wt or mutated (phospho-mutant S824A or phospho-mimic S824D) KAP-1 plasmids. γH2AX foci were analyzed in GFP-positive G2-irradiated cells. EdU and cell cycle markers were used to distinguish G2- from S- and G1-phase cells . In (A), (B) and (C), foci numbers or PCC breaks from unirradiated cells were subtracted. At least 40 cells or PCC spreads were analyzed per data point and experiment (mean ± SEM from ≥3 experiments). KAP-1 and BRCA2 depletion in this and subsequent experiments was highly efficient (>90% as assessed by Western blotting). P values were obtained by t -test and represent a comparison of all cells analyzed in the indicated cell populations (***: p<0.001).
    Figure Legend Snippet: ( A ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) and HSC62 hTert (BRCA2-deficient) human fibroblasts. ( B ) RPA and RAD51 foci analysis in G2-irradiated A549 tumor cells. ( C ) Endogenous KAP-1 and BRCA2 was depleted in HeLa tumor cells by siRNA, and cells were transfected with GFP-tagged and siRNA-resistant empty (GFP), wt or mutated (phospho-mutant S824A or phospho-mimic S824D) KAP-1 plasmids. γH2AX foci were analyzed in GFP-positive G2-irradiated cells. EdU and cell cycle markers were used to distinguish G2- from S- and G1-phase cells . In (A), (B) and (C), foci numbers or PCC breaks from unirradiated cells were subtracted. At least 40 cells or PCC spreads were analyzed per data point and experiment (mean ± SEM from ≥3 experiments). KAP-1 and BRCA2 depletion in this and subsequent experiments was highly efficient (>90% as assessed by Western blotting). P values were obtained by t -test and represent a comparison of all cells analyzed in the indicated cell populations (***: p<0.001).

    Techniques Used: Irradiation, Transfection, Mutagenesis, Western Blot, Comparison

    ( A ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) and 2BN hTert (XLF-deficient) human fibroblasts. ( B ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) human fibroblasts treated with PARPi 0.5 h prior to IR. ( C ) PCC analysis from G2-irradiated HSC62 hTert (BRCA2-deficient) human fibroblasts treated with PARPi as in (B). ( D ) γH2AX foci analysis in G2-irradiated 82-6 hTert (wt) and HSC62 hTert (BRCA2-deficient) human fibroblasts. ( E–G ) Chromatid fusions and breaks in G2-irradiated mitotic HeLa tumor cells at 8 h post 2 Gy. Cells were treated with caffeine and colcemid at 5 h post IR to abolish the G2 checkpoint and collected in mitosis. Chromosomes were stained with Giemsa (panel E) or analyzed by FISH with probes specific for chromosomes 1 (red), 2 (green) and 4 (yellow) (panel F) or to chromosomes 18 and 19 (panel G). Foci numbers, chromatid breaks and fusions from unirradiated cells were subtracted. For panels A–E, at least 40 cells or 40 PCC/mitotic spreads were analyzed per data point and experiment (mean ± SEM from ≥3 experiments). For panel G, 50 mitotic spreads were analyzed per data point and experiment (mean ± SEM from ≥2 experiments). N.d. indicates that no chromatid fusions were observed under these conditions. P values were obtained by t -test and represent a comparison of all cells analyzed in the indicated cell populations (***: p<0.001).
    Figure Legend Snippet: ( A ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) and 2BN hTert (XLF-deficient) human fibroblasts. ( B ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) human fibroblasts treated with PARPi 0.5 h prior to IR. ( C ) PCC analysis from G2-irradiated HSC62 hTert (BRCA2-deficient) human fibroblasts treated with PARPi as in (B). ( D ) γH2AX foci analysis in G2-irradiated 82-6 hTert (wt) and HSC62 hTert (BRCA2-deficient) human fibroblasts. ( E–G ) Chromatid fusions and breaks in G2-irradiated mitotic HeLa tumor cells at 8 h post 2 Gy. Cells were treated with caffeine and colcemid at 5 h post IR to abolish the G2 checkpoint and collected in mitosis. Chromosomes were stained with Giemsa (panel E) or analyzed by FISH with probes specific for chromosomes 1 (red), 2 (green) and 4 (yellow) (panel F) or to chromosomes 18 and 19 (panel G). Foci numbers, chromatid breaks and fusions from unirradiated cells were subtracted. For panels A–E, at least 40 cells or 40 PCC/mitotic spreads were analyzed per data point and experiment (mean ± SEM from ≥3 experiments). For panel G, 50 mitotic spreads were analyzed per data point and experiment (mean ± SEM from ≥2 experiments). N.d. indicates that no chromatid fusions were observed under these conditions. P values were obtained by t -test and represent a comparison of all cells analyzed in the indicated cell populations (***: p<0.001).

    Techniques Used: Irradiation, Staining, Comparison



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    ( A ) A549 tumor cells treated with <t>BRCA2</t> siRNA were irradiated with 1 Gy (0.5 h) or 2 Gy (8 h) and immunostained with the indicated antibodies. Using EdU and cell cycle markers to distinguish G1- from G2-phase cells , focal intensities of pATM were measured using ImageJ software (see ). BRCA2 siRNA was used in this analysis to accumulate resected DSBs. ( B ) 2BN hTert (XLF-deficient) human fibroblasts were analyzed 2 h post IR with 1 Gy. Cells were stained against γH2AX and RAD51 or pATM and RAD51, and γH2AX or pATM focal intensities were measured at RAD51-foci-positive or RAD51-foci-negative foci. XLF-deficient cells were used in this analysis to prevent repair of EC DSBs during the time needed for resection of HC DSBs. ( C ) 82-6 hTert (wt) and F02-98 hTert (ATR-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). ( D ) 2BN hTert (XLF-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). Since both DNA-PK and ATM can phosphorylate γH2AX, cells were treated with DNA-PK inhibitor under all conditions. Inhibitors were added 1 h post IR, a time sufficient to allow for ATM-dependent resection and RAD51 loading (see ). In (A–D), at least 300 foci from 3 independent experiments were analyzed for each point. Box plots were used with a maximum whisker-length of 1.5-fold the inter-quartile range; the lower and upper “x” indicates the 1% or 99% margin of the data range, respectively.
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    Image Search Results


    ( A ) A549 tumor cells treated with BRCA2 siRNA were irradiated with 1 Gy (0.5 h) or 2 Gy (8 h) and immunostained with the indicated antibodies. Using EdU and cell cycle markers to distinguish G1- from G2-phase cells , focal intensities of pATM were measured using ImageJ software (see ). BRCA2 siRNA was used in this analysis to accumulate resected DSBs. ( B ) 2BN hTert (XLF-deficient) human fibroblasts were analyzed 2 h post IR with 1 Gy. Cells were stained against γH2AX and RAD51 or pATM and RAD51, and γH2AX or pATM focal intensities were measured at RAD51-foci-positive or RAD51-foci-negative foci. XLF-deficient cells were used in this analysis to prevent repair of EC DSBs during the time needed for resection of HC DSBs. ( C ) 82-6 hTert (wt) and F02-98 hTert (ATR-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). ( D ) 2BN hTert (XLF-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). Since both DNA-PK and ATM can phosphorylate γH2AX, cells were treated with DNA-PK inhibitor under all conditions. Inhibitors were added 1 h post IR, a time sufficient to allow for ATM-dependent resection and RAD51 loading (see ). In (A–D), at least 300 foci from 3 independent experiments were analyzed for each point. Box plots were used with a maximum whisker-length of 1.5-fold the inter-quartile range; the lower and upper “x” indicates the 1% or 99% margin of the data range, respectively.

    Journal: PLoS Genetics

    Article Title: ATM Release at Resected Double-Strand Breaks Provides Heterochromatin Reconstitution to Facilitate Homologous Recombination

    doi: 10.1371/journal.pgen.1003667

    Figure Lengend Snippet: ( A ) A549 tumor cells treated with BRCA2 siRNA were irradiated with 1 Gy (0.5 h) or 2 Gy (8 h) and immunostained with the indicated antibodies. Using EdU and cell cycle markers to distinguish G1- from G2-phase cells , focal intensities of pATM were measured using ImageJ software (see ). BRCA2 siRNA was used in this analysis to accumulate resected DSBs. ( B ) 2BN hTert (XLF-deficient) human fibroblasts were analyzed 2 h post IR with 1 Gy. Cells were stained against γH2AX and RAD51 or pATM and RAD51, and γH2AX or pATM focal intensities were measured at RAD51-foci-positive or RAD51-foci-negative foci. XLF-deficient cells were used in this analysis to prevent repair of EC DSBs during the time needed for resection of HC DSBs. ( C ) 82-6 hTert (wt) and F02-98 hTert (ATR-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). ( D ) 2BN hTert (XLF-deficient) human fibroblasts were stained against γH2AX and RAD51 at 2 h post 1 Gy, and γH2AX focal intensities were measured as in (B). Since both DNA-PK and ATM can phosphorylate γH2AX, cells were treated with DNA-PK inhibitor under all conditions. Inhibitors were added 1 h post IR, a time sufficient to allow for ATM-dependent resection and RAD51 loading (see ). In (A–D), at least 300 foci from 3 independent experiments were analyzed for each point. Box plots were used with a maximum whisker-length of 1.5-fold the inter-quartile range; the lower and upper “x” indicates the 1% or 99% margin of the data range, respectively.

    Article Snippet: Primary antibodies used were: rabbit-α-pATM at 1∶1000 (Epitomics); rabbit-α-pKAP-1 (S824) at 1∶10000 (Epitomics); rabbit-α-KAP-1 at 1∶1000 (abcam); mouse-α-BRCA2 at 1∶1000 (Cell signaling); rabbit-α-GAPDH at 1∶1000 (Santa Cruz); mouse-α-γH2AX at 1∶1000 (Millipore); mouse-α-H3 at 1∶1000 (abcam); mouse-α-RPA2 at 1∶1000 (Calbiochem); rabbit-α-pRPA2 (S4/8) at 1∶10000 (Bethyl).

    Techniques: Irradiation, Software, Staining, Whisker Assay

    ( A ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) and HSC62 hTert (BRCA2-deficient) human fibroblasts. ( B ) RPA and RAD51 foci analysis in G2-irradiated A549 tumor cells. ( C ) Endogenous KAP-1 and BRCA2 was depleted in HeLa tumor cells by siRNA, and cells were transfected with GFP-tagged and siRNA-resistant empty (GFP), wt or mutated (phospho-mutant S824A or phospho-mimic S824D) KAP-1 plasmids. γH2AX foci were analyzed in GFP-positive G2-irradiated cells. EdU and cell cycle markers were used to distinguish G2- from S- and G1-phase cells . In (A), (B) and (C), foci numbers or PCC breaks from unirradiated cells were subtracted. At least 40 cells or PCC spreads were analyzed per data point and experiment (mean ± SEM from ≥3 experiments). KAP-1 and BRCA2 depletion in this and subsequent experiments was highly efficient (>90% as assessed by Western blotting). P values were obtained by t -test and represent a comparison of all cells analyzed in the indicated cell populations (***: p<0.001).

    Journal: PLoS Genetics

    Article Title: ATM Release at Resected Double-Strand Breaks Provides Heterochromatin Reconstitution to Facilitate Homologous Recombination

    doi: 10.1371/journal.pgen.1003667

    Figure Lengend Snippet: ( A ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) and HSC62 hTert (BRCA2-deficient) human fibroblasts. ( B ) RPA and RAD51 foci analysis in G2-irradiated A549 tumor cells. ( C ) Endogenous KAP-1 and BRCA2 was depleted in HeLa tumor cells by siRNA, and cells were transfected with GFP-tagged and siRNA-resistant empty (GFP), wt or mutated (phospho-mutant S824A or phospho-mimic S824D) KAP-1 plasmids. γH2AX foci were analyzed in GFP-positive G2-irradiated cells. EdU and cell cycle markers were used to distinguish G2- from S- and G1-phase cells . In (A), (B) and (C), foci numbers or PCC breaks from unirradiated cells were subtracted. At least 40 cells or PCC spreads were analyzed per data point and experiment (mean ± SEM from ≥3 experiments). KAP-1 and BRCA2 depletion in this and subsequent experiments was highly efficient (>90% as assessed by Western blotting). P values were obtained by t -test and represent a comparison of all cells analyzed in the indicated cell populations (***: p<0.001).

    Article Snippet: Primary antibodies used were: rabbit-α-pATM at 1∶1000 (Epitomics); rabbit-α-pKAP-1 (S824) at 1∶10000 (Epitomics); rabbit-α-KAP-1 at 1∶1000 (abcam); mouse-α-BRCA2 at 1∶1000 (Cell signaling); rabbit-α-GAPDH at 1∶1000 (Santa Cruz); mouse-α-γH2AX at 1∶1000 (Millipore); mouse-α-H3 at 1∶1000 (abcam); mouse-α-RPA2 at 1∶1000 (Calbiochem); rabbit-α-pRPA2 (S4/8) at 1∶10000 (Bethyl).

    Techniques: Irradiation, Transfection, Mutagenesis, Western Blot, Comparison

    ( A ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) and 2BN hTert (XLF-deficient) human fibroblasts. ( B ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) human fibroblasts treated with PARPi 0.5 h prior to IR. ( C ) PCC analysis from G2-irradiated HSC62 hTert (BRCA2-deficient) human fibroblasts treated with PARPi as in (B). ( D ) γH2AX foci analysis in G2-irradiated 82-6 hTert (wt) and HSC62 hTert (BRCA2-deficient) human fibroblasts. ( E–G ) Chromatid fusions and breaks in G2-irradiated mitotic HeLa tumor cells at 8 h post 2 Gy. Cells were treated with caffeine and colcemid at 5 h post IR to abolish the G2 checkpoint and collected in mitosis. Chromosomes were stained with Giemsa (panel E) or analyzed by FISH with probes specific for chromosomes 1 (red), 2 (green) and 4 (yellow) (panel F) or to chromosomes 18 and 19 (panel G). Foci numbers, chromatid breaks and fusions from unirradiated cells were subtracted. For panels A–E, at least 40 cells or 40 PCC/mitotic spreads were analyzed per data point and experiment (mean ± SEM from ≥3 experiments). For panel G, 50 mitotic spreads were analyzed per data point and experiment (mean ± SEM from ≥2 experiments). N.d. indicates that no chromatid fusions were observed under these conditions. P values were obtained by t -test and represent a comparison of all cells analyzed in the indicated cell populations (***: p<0.001).

    Journal: PLoS Genetics

    Article Title: ATM Release at Resected Double-Strand Breaks Provides Heterochromatin Reconstitution to Facilitate Homologous Recombination

    doi: 10.1371/journal.pgen.1003667

    Figure Lengend Snippet: ( A ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) and 2BN hTert (XLF-deficient) human fibroblasts. ( B ) γH2AX foci and PCC analysis in G2-irradiated 82-6 hTert (wt) human fibroblasts treated with PARPi 0.5 h prior to IR. ( C ) PCC analysis from G2-irradiated HSC62 hTert (BRCA2-deficient) human fibroblasts treated with PARPi as in (B). ( D ) γH2AX foci analysis in G2-irradiated 82-6 hTert (wt) and HSC62 hTert (BRCA2-deficient) human fibroblasts. ( E–G ) Chromatid fusions and breaks in G2-irradiated mitotic HeLa tumor cells at 8 h post 2 Gy. Cells were treated with caffeine and colcemid at 5 h post IR to abolish the G2 checkpoint and collected in mitosis. Chromosomes were stained with Giemsa (panel E) or analyzed by FISH with probes specific for chromosomes 1 (red), 2 (green) and 4 (yellow) (panel F) or to chromosomes 18 and 19 (panel G). Foci numbers, chromatid breaks and fusions from unirradiated cells were subtracted. For panels A–E, at least 40 cells or 40 PCC/mitotic spreads were analyzed per data point and experiment (mean ± SEM from ≥3 experiments). For panel G, 50 mitotic spreads were analyzed per data point and experiment (mean ± SEM from ≥2 experiments). N.d. indicates that no chromatid fusions were observed under these conditions. P values were obtained by t -test and represent a comparison of all cells analyzed in the indicated cell populations (***: p<0.001).

    Article Snippet: Primary antibodies used were: rabbit-α-pATM at 1∶1000 (Epitomics); rabbit-α-pKAP-1 (S824) at 1∶10000 (Epitomics); rabbit-α-KAP-1 at 1∶1000 (abcam); mouse-α-BRCA2 at 1∶1000 (Cell signaling); rabbit-α-GAPDH at 1∶1000 (Santa Cruz); mouse-α-γH2AX at 1∶1000 (Millipore); mouse-α-H3 at 1∶1000 (abcam); mouse-α-RPA2 at 1∶1000 (Calbiochem); rabbit-α-pRPA2 (S4/8) at 1∶10000 (Bethyl).

    Techniques: Irradiation, Staining, Comparison