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  • 93
    Carl Zeiss axio imager a1 microscope
    Effects of anti-HER2 antibody BH1 and tumor necrosis factor-α on DNA synthesis and cell cycle in SK-BR-3 cells. ( A ) Effects on DNA synthesis were tested by 24-h BrdU incorporation assay. Cells were plated on coverslips in 12-well plates, treated as described in Figure 5 , and were exposed to BrdU during 24 h prior to indicated days. Nuclear BrdU was detected by indirect immunofluorescence assay. Nuclei were counterstained with DAPI, examined using Zeiss <t>Axio</t> Imager.A1 microscope. Representative photographs (3–5 frames per coverslip) were acquired. Percent BrdU incorporation was calculated after manual counting of BrdU + and BrdU - nuclei. * Denotes p-values ≤ 0.05 (BH1 vs. IgG1, BH1 + TNF-α vs. IgG1 + TNF-α). T-bars: SD. ( B - D ). Effects on cell cycle were analyzed by flow cytometry. SK-BR-3 cells were treated with 20 μg/ml BH1 or IgG isotype control antibody in the presence or absence of 1000 U/ml TNF-α. Cells were fixed on day 1 ( B ), day 3 ( C ) and day 6 ( D ), stained with propidium iodide and the cell cycle distribution was analysed with flow cytometry. Fraction of cells present in G2/M-phase (G2/M), S-phase (S) and G1-phase (G1) are shown in purple, green and orange respectively. Fraction of apoptotic cells observed as a SubG1 peak is shown in black.
    Axio Imager A1 Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 93/100, based on 1863 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Carl Zeiss axio imager a1 fluorescence microscope
    Effects of anti-HER2 antibody BH1 and tumor necrosis factor-α on DNA synthesis and cell cycle in SK-BR-3 cells. ( A ) Effects on DNA synthesis were tested by 24-h BrdU incorporation assay. Cells were plated on coverslips in 12-well plates, treated as described in Figure 5 , and were exposed to BrdU during 24 h prior to indicated days. Nuclear BrdU was detected by indirect immunofluorescence assay. Nuclei were counterstained with DAPI, examined using Zeiss <t>Axio</t> Imager.A1 microscope. Representative photographs (3–5 frames per coverslip) were acquired. Percent BrdU incorporation was calculated after manual counting of BrdU + and BrdU - nuclei. * Denotes p-values ≤ 0.05 (BH1 vs. IgG1, BH1 + TNF-α vs. IgG1 + TNF-α). T-bars: SD. ( B - D ). Effects on cell cycle were analyzed by flow cytometry. SK-BR-3 cells were treated with 20 μg/ml BH1 or IgG isotype control antibody in the presence or absence of 1000 U/ml TNF-α. Cells were fixed on day 1 ( B ), day 3 ( C ) and day 6 ( D ), stained with propidium iodide and the cell cycle distribution was analysed with flow cytometry. Fraction of cells present in G2/M-phase (G2/M), S-phase (S) and G1-phase (G1) are shown in purple, green and orange respectively. Fraction of apoptotic cells observed as a SubG1 peak is shown in black.
    Axio Imager A1 Fluorescence Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 91/100, based on 498 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Carl Zeiss axioimager a1 fluorescent microscope
    Effects of anti-HER2 antibody BH1 and tumor necrosis factor-α on DNA synthesis and cell cycle in SK-BR-3 cells. ( A ) Effects on DNA synthesis were tested by 24-h BrdU incorporation assay. Cells were plated on coverslips in 12-well plates, treated as described in Figure 5 , and were exposed to BrdU during 24 h prior to indicated days. Nuclear BrdU was detected by indirect immunofluorescence assay. Nuclei were counterstained with DAPI, examined using Zeiss <t>Axio</t> Imager.A1 microscope. Representative photographs (3–5 frames per coverslip) were acquired. Percent BrdU incorporation was calculated after manual counting of BrdU + and BrdU - nuclei. * Denotes p-values ≤ 0.05 (BH1 vs. IgG1, BH1 + TNF-α vs. IgG1 + TNF-α). T-bars: SD. ( B - D ). Effects on cell cycle were analyzed by flow cytometry. SK-BR-3 cells were treated with 20 μg/ml BH1 or IgG isotype control antibody in the presence or absence of 1000 U/ml TNF-α. Cells were fixed on day 1 ( B ), day 3 ( C ) and day 6 ( D ), stained with propidium iodide and the cell cycle distribution was analysed with flow cytometry. Fraction of cells present in G2/M-phase (G2/M), S-phase (S) and G1-phase (G1) are shown in purple, green and orange respectively. Fraction of apoptotic cells observed as a SubG1 peak is shown in black.
    Axioimager A1 Fluorescent Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 88/100, based on 142 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Carl Zeiss axio imager a1 epifluorescence microscope
    Effects of anti-HER2 antibody BH1 and tumor necrosis factor-α on DNA synthesis and cell cycle in SK-BR-3 cells. ( A ) Effects on DNA synthesis were tested by 24-h BrdU incorporation assay. Cells were plated on coverslips in 12-well plates, treated as described in Figure 5 , and were exposed to BrdU during 24 h prior to indicated days. Nuclear BrdU was detected by indirect immunofluorescence assay. Nuclei were counterstained with DAPI, examined using Zeiss <t>Axio</t> Imager.A1 microscope. Representative photographs (3–5 frames per coverslip) were acquired. Percent BrdU incorporation was calculated after manual counting of BrdU + and BrdU - nuclei. * Denotes p-values ≤ 0.05 (BH1 vs. IgG1, BH1 + TNF-α vs. IgG1 + TNF-α). T-bars: SD. ( B - D ). Effects on cell cycle were analyzed by flow cytometry. SK-BR-3 cells were treated with 20 μg/ml BH1 or IgG isotype control antibody in the presence or absence of 1000 U/ml TNF-α. Cells were fixed on day 1 ( B ), day 3 ( C ) and day 6 ( D ), stained with propidium iodide and the cell cycle distribution was analysed with flow cytometry. Fraction of cells present in G2/M-phase (G2/M), S-phase (S) and G1-phase (G1) are shown in purple, green and orange respectively. Fraction of apoptotic cells observed as a SubG1 peak is shown in black.
    Axio Imager A1 Epifluorescence Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 89/100, based on 104 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/axio imager a1 epifluorescence microscope/product/Carl Zeiss
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    Price from $9.99 to $1999.99
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    88
    Carl Zeiss axio imager a 1 microscope
    Effects of anti-HER2 antibody BH1 and tumor necrosis factor-α on DNA synthesis and cell cycle in SK-BR-3 cells. ( A ) Effects on DNA synthesis were tested by 24-h BrdU incorporation assay. Cells were plated on coverslips in 12-well plates, treated as described in Figure 5 , and were exposed to BrdU during 24 h prior to indicated days. Nuclear BrdU was detected by indirect immunofluorescence assay. Nuclei were counterstained with DAPI, examined using Zeiss <t>Axio</t> Imager.A1 microscope. Representative photographs (3–5 frames per coverslip) were acquired. Percent BrdU incorporation was calculated after manual counting of BrdU + and BrdU - nuclei. * Denotes p-values ≤ 0.05 (BH1 vs. IgG1, BH1 + TNF-α vs. IgG1 + TNF-α). T-bars: SD. ( B - D ). Effects on cell cycle were analyzed by flow cytometry. SK-BR-3 cells were treated with 20 μg/ml BH1 or IgG isotype control antibody in the presence or absence of 1000 U/ml TNF-α. Cells were fixed on day 1 ( B ), day 3 ( C ) and day 6 ( D ), stained with propidium iodide and the cell cycle distribution was analysed with flow cytometry. Fraction of cells present in G2/M-phase (G2/M), S-phase (S) and G1-phase (G1) are shown in purple, green and orange respectively. Fraction of apoptotic cells observed as a SubG1 peak is shown in black.
    Axio Imager A 1 Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 88/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/axio imager a 1 microscope/product/Carl Zeiss
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    92
    Carl Zeiss axio imager a1 compound microscope
    Effects of anti-HER2 antibody BH1 and tumor necrosis factor-α on DNA synthesis and cell cycle in SK-BR-3 cells. ( A ) Effects on DNA synthesis were tested by 24-h BrdU incorporation assay. Cells were plated on coverslips in 12-well plates, treated as described in Figure 5 , and were exposed to BrdU during 24 h prior to indicated days. Nuclear BrdU was detected by indirect immunofluorescence assay. Nuclei were counterstained with DAPI, examined using Zeiss <t>Axio</t> Imager.A1 microscope. Representative photographs (3–5 frames per coverslip) were acquired. Percent BrdU incorporation was calculated after manual counting of BrdU + and BrdU - nuclei. * Denotes p-values ≤ 0.05 (BH1 vs. IgG1, BH1 + TNF-α vs. IgG1 + TNF-α). T-bars: SD. ( B - D ). Effects on cell cycle were analyzed by flow cytometry. SK-BR-3 cells were treated with 20 μg/ml BH1 or IgG isotype control antibody in the presence or absence of 1000 U/ml TNF-α. Cells were fixed on day 1 ( B ), day 3 ( C ) and day 6 ( D ), stained with propidium iodide and the cell cycle distribution was analysed with flow cytometry. Fraction of cells present in G2/M-phase (G2/M), S-phase (S) and G1-phase (G1) are shown in purple, green and orange respectively. Fraction of apoptotic cells observed as a SubG1 peak is shown in black.
    Axio Imager A1 Compound Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 92/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Carl Zeiss axioimager a1 upright microscope
    Effects of anti-HER2 antibody BH1 and tumor necrosis factor-α on DNA synthesis and cell cycle in SK-BR-3 cells. ( A ) Effects on DNA synthesis were tested by 24-h BrdU incorporation assay. Cells were plated on coverslips in 12-well plates, treated as described in Figure 5 , and were exposed to BrdU during 24 h prior to indicated days. Nuclear BrdU was detected by indirect immunofluorescence assay. Nuclei were counterstained with DAPI, examined using Zeiss <t>Axio</t> Imager.A1 microscope. Representative photographs (3–5 frames per coverslip) were acquired. Percent BrdU incorporation was calculated after manual counting of BrdU + and BrdU - nuclei. * Denotes p-values ≤ 0.05 (BH1 vs. IgG1, BH1 + TNF-α vs. IgG1 + TNF-α). T-bars: SD. ( B - D ). Effects on cell cycle were analyzed by flow cytometry. SK-BR-3 cells were treated with 20 μg/ml BH1 or IgG isotype control antibody in the presence or absence of 1000 U/ml TNF-α. Cells were fixed on day 1 ( B ), day 3 ( C ) and day 6 ( D ), stained with propidium iodide and the cell cycle distribution was analysed with flow cytometry. Fraction of cells present in G2/M-phase (G2/M), S-phase (S) and G1-phase (G1) are shown in purple, green and orange respectively. Fraction of apoptotic cells observed as a SubG1 peak is shown in black.
    Axioimager A1 Upright Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 89/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Carl Zeiss axioimager a1 upright epifluorescent microscope
    The FANCD2 NLS mutants fail to correct the ICL sensitivity of FA-D2 patient cells. ( A ) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100, FANCD2-3N, or FANCD2-K561R were incubated in the absence or presence of 8 or 16 nM MMC for 24 h and the numbers of chromosome aberrations including gaps and breaks, dicentrics, and complex chromosome aberrations, including radial formations, were scored. Metaphase spreads were analyzed using a Zeiss <t>AxioImager.A1</t> upright <t>epifluorescent</t> microscope with AxioVision LE 4.6 image acquisition software. At least 80 metaphases were scored per treatment. Error bars represent the standard error of the means. ( B ) FA-D2 cells stably expressing LacZ, FANCD2-WT and FANCD2-∆N57 were incubated in the absence (NT) or presence of 40 nM MMC for 18 h, and allowed to recover for 0, 4.5 or 7 h. Cells were then fixed, stained with rabbit polyclonal anti-DNA-PK CS pS2056, and counterstained with DAPI. At least 300 cells were scored for nuclei with > 5 DNA-PK CS foci. Error bars represent the standard error of the means from two independent experiments. ***, p
    Axioimager A1 Upright Epifluorescent Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 88/100, based on 63 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/axioimager a1 upright epifluorescent microscope/product/Carl Zeiss
    Average 88 stars, based on 63 article reviews
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    92
    Carl Zeiss axio imager a 1
    The FANCD2 NLS mutants fail to correct the ICL sensitivity of FA-D2 patient cells. ( A ) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100, FANCD2-3N, or FANCD2-K561R were incubated in the absence or presence of 8 or 16 nM MMC for 24 h and the numbers of chromosome aberrations including gaps and breaks, dicentrics, and complex chromosome aberrations, including radial formations, were scored. Metaphase spreads were analyzed using a Zeiss <t>AxioImager.A1</t> upright <t>epifluorescent</t> microscope with AxioVision LE 4.6 image acquisition software. At least 80 metaphases were scored per treatment. Error bars represent the standard error of the means. ( B ) FA-D2 cells stably expressing LacZ, FANCD2-WT and FANCD2-∆N57 were incubated in the absence (NT) or presence of 40 nM MMC for 18 h, and allowed to recover for 0, 4.5 or 7 h. Cells were then fixed, stained with rabbit polyclonal anti-DNA-PK CS pS2056, and counterstained with DAPI. At least 300 cells were scored for nuclei with > 5 DNA-PK CS foci. Error bars represent the standard error of the means from two independent experiments. ***, p
    Axio Imager A 1, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 92/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/axio imager a 1/product/Carl Zeiss
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    axio imager a 1 - by Bioz Stars, 2020-10
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    86
    Carl Zeiss axio imager a1 epifluorescent microscope
    The FANCD2 NLS mutants fail to correct the ICL sensitivity of FA-D2 patient cells. ( A ) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100, FANCD2-3N, or FANCD2-K561R were incubated in the absence or presence of 8 or 16 nM MMC for 24 h and the numbers of chromosome aberrations including gaps and breaks, dicentrics, and complex chromosome aberrations, including radial formations, were scored. Metaphase spreads were analyzed using a Zeiss <t>AxioImager.A1</t> upright <t>epifluorescent</t> microscope with AxioVision LE 4.6 image acquisition software. At least 80 metaphases were scored per treatment. Error bars represent the standard error of the means. ( B ) FA-D2 cells stably expressing LacZ, FANCD2-WT and FANCD2-∆N57 were incubated in the absence (NT) or presence of 40 nM MMC for 18 h, and allowed to recover for 0, 4.5 or 7 h. Cells were then fixed, stained with rabbit polyclonal anti-DNA-PK CS pS2056, and counterstained with DAPI. At least 300 cells were scored for nuclei with > 5 DNA-PK CS foci. Error bars represent the standard error of the means from two independent experiments. ***, p
    Axio Imager A1 Epifluorescent Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 86/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/axio imager a1 epifluorescent microscope/product/Carl Zeiss
    Average 86 stars, based on 31 article reviews
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    axio imager a1 epifluorescent microscope - by Bioz Stars, 2020-10
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    90
    Carl Zeiss axioimager a1 upright epifluorescence microscope
    The FANCD2 NLS mutants fail to correct the ICL sensitivity of FA-D2 patient cells. ( A ) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100, FANCD2-3N, or FANCD2-K561R were incubated in the absence or presence of 8 or 16 nM MMC for 24 h and the numbers of chromosome aberrations including gaps and breaks, dicentrics, and complex chromosome aberrations, including radial formations, were scored. Metaphase spreads were analyzed using a Zeiss <t>AxioImager.A1</t> upright <t>epifluorescent</t> microscope with AxioVision LE 4.6 image acquisition software. At least 80 metaphases were scored per treatment. Error bars represent the standard error of the means. ( B ) FA-D2 cells stably expressing LacZ, FANCD2-WT and FANCD2-∆N57 were incubated in the absence (NT) or presence of 40 nM MMC for 18 h, and allowed to recover for 0, 4.5 or 7 h. Cells were then fixed, stained with rabbit polyclonal anti-DNA-PK CS pS2056, and counterstained with DAPI. At least 300 cells were scored for nuclei with > 5 DNA-PK CS foci. Error bars represent the standard error of the means from two independent experiments. ***, p
    Axioimager A1 Upright Epifluorescence Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 90/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/axioimager a1 upright epifluorescence microscope/product/Carl Zeiss
    Average 90 stars, based on 20 article reviews
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    Image Search Results


    Effects of anti-HER2 antibody BH1 and tumor necrosis factor-α on DNA synthesis and cell cycle in SK-BR-3 cells. ( A ) Effects on DNA synthesis were tested by 24-h BrdU incorporation assay. Cells were plated on coverslips in 12-well plates, treated as described in Figure 5 , and were exposed to BrdU during 24 h prior to indicated days. Nuclear BrdU was detected by indirect immunofluorescence assay. Nuclei were counterstained with DAPI, examined using Zeiss Axio Imager.A1 microscope. Representative photographs (3–5 frames per coverslip) were acquired. Percent BrdU incorporation was calculated after manual counting of BrdU + and BrdU - nuclei. * Denotes p-values ≤ 0.05 (BH1 vs. IgG1, BH1 + TNF-α vs. IgG1 + TNF-α). T-bars: SD. ( B - D ). Effects on cell cycle were analyzed by flow cytometry. SK-BR-3 cells were treated with 20 μg/ml BH1 or IgG isotype control antibody in the presence or absence of 1000 U/ml TNF-α. Cells were fixed on day 1 ( B ), day 3 ( C ) and day 6 ( D ), stained with propidium iodide and the cell cycle distribution was analysed with flow cytometry. Fraction of cells present in G2/M-phase (G2/M), S-phase (S) and G1-phase (G1) are shown in purple, green and orange respectively. Fraction of apoptotic cells observed as a SubG1 peak is shown in black.

    Journal: BMC Cancer

    Article Title: Novel anti-HER2 monoclonal antibodies: synergy and antagonism with tumor necrosis factor-?

    doi: 10.1186/1471-2407-12-450

    Figure Lengend Snippet: Effects of anti-HER2 antibody BH1 and tumor necrosis factor-α on DNA synthesis and cell cycle in SK-BR-3 cells. ( A ) Effects on DNA synthesis were tested by 24-h BrdU incorporation assay. Cells were plated on coverslips in 12-well plates, treated as described in Figure 5 , and were exposed to BrdU during 24 h prior to indicated days. Nuclear BrdU was detected by indirect immunofluorescence assay. Nuclei were counterstained with DAPI, examined using Zeiss Axio Imager.A1 microscope. Representative photographs (3–5 frames per coverslip) were acquired. Percent BrdU incorporation was calculated after manual counting of BrdU + and BrdU - nuclei. * Denotes p-values ≤ 0.05 (BH1 vs. IgG1, BH1 + TNF-α vs. IgG1 + TNF-α). T-bars: SD. ( B - D ). Effects on cell cycle were analyzed by flow cytometry. SK-BR-3 cells were treated with 20 μg/ml BH1 or IgG isotype control antibody in the presence or absence of 1000 U/ml TNF-α. Cells were fixed on day 1 ( B ), day 3 ( C ) and day 6 ( D ), stained with propidium iodide and the cell cycle distribution was analysed with flow cytometry. Fraction of cells present in G2/M-phase (G2/M), S-phase (S) and G1-phase (G1) are shown in purple, green and orange respectively. Fraction of apoptotic cells observed as a SubG1 peak is shown in black.

    Article Snippet: Nuclei were counterstained with DAPI and cells were observed using Zeiss Axio Imager.A1 microscope (Jena, Germany).

    Techniques: DNA Synthesis, BrdU Incorporation Assay, Immunofluorescence, Microscopy, Flow Cytometry, Cytometry, Staining

    Strong binding of anti-HER2 antibodies to ERBB2 -amplified breast cancer cell lines. ERRB2 amplification was analyzed by FISH analysis. Cells were harvested at confluence, fixed and subjected to dual-color FISH using Texas Red-labeled ERBB2 (red) and FITC-labeled chromosome 17 α-satellite CEP17 (green) DNA probes (left panel). For generation of anti-HER2 monoclonal antibodies, mice were immunized with SK-BR-3 cells, followed by a recombinant protein composed of extracellular domain of HER2 fused to human IgG1 Fc domain (HER2 ECD). Antibodies were purified from conditioned hybridoma cell culture media by affinity chromatography. For immunofluorescence assay, cells were seeded on coverslips in 6-well plates, incubated 36 h in cell culture medium, fixed, and permeabilized. Fixed cells were incubated with anti-HER2 antibodies, and the primary antibody binding was detected using Alexa 488- conjugated anti-mouse IgG (BH1, BH2, BH5, BH6, BH7 panels; green). Nuclei were counterstained with DAPI (blue) and pictures were acquired using Zeiss Axio Imager.A1 microscope.

    Journal: BMC Cancer

    Article Title: Novel anti-HER2 monoclonal antibodies: synergy and antagonism with tumor necrosis factor-?

    doi: 10.1186/1471-2407-12-450

    Figure Lengend Snippet: Strong binding of anti-HER2 antibodies to ERBB2 -amplified breast cancer cell lines. ERRB2 amplification was analyzed by FISH analysis. Cells were harvested at confluence, fixed and subjected to dual-color FISH using Texas Red-labeled ERBB2 (red) and FITC-labeled chromosome 17 α-satellite CEP17 (green) DNA probes (left panel). For generation of anti-HER2 monoclonal antibodies, mice were immunized with SK-BR-3 cells, followed by a recombinant protein composed of extracellular domain of HER2 fused to human IgG1 Fc domain (HER2 ECD). Antibodies were purified from conditioned hybridoma cell culture media by affinity chromatography. For immunofluorescence assay, cells were seeded on coverslips in 6-well plates, incubated 36 h in cell culture medium, fixed, and permeabilized. Fixed cells were incubated with anti-HER2 antibodies, and the primary antibody binding was detected using Alexa 488- conjugated anti-mouse IgG (BH1, BH2, BH5, BH6, BH7 panels; green). Nuclei were counterstained with DAPI (blue) and pictures were acquired using Zeiss Axio Imager.A1 microscope.

    Article Snippet: Nuclei were counterstained with DAPI and cells were observed using Zeiss Axio Imager.A1 microscope (Jena, Germany).

    Techniques: Binding Assay, Amplification, Fluorescence In Situ Hybridization, Labeling, Mouse Assay, Recombinant, Purification, Cell Culture, Affinity Chromatography, Immunofluorescence, Incubation, Microscopy

    The FANCD2 NLS mutants fail to correct the ICL sensitivity of FA-D2 patient cells. ( A ) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100, FANCD2-3N, or FANCD2-K561R were incubated in the absence or presence of 8 or 16 nM MMC for 24 h and the numbers of chromosome aberrations including gaps and breaks, dicentrics, and complex chromosome aberrations, including radial formations, were scored. Metaphase spreads were analyzed using a Zeiss AxioImager.A1 upright epifluorescent microscope with AxioVision LE 4.6 image acquisition software. At least 80 metaphases were scored per treatment. Error bars represent the standard error of the means. ( B ) FA-D2 cells stably expressing LacZ, FANCD2-WT and FANCD2-∆N57 were incubated in the absence (NT) or presence of 40 nM MMC for 18 h, and allowed to recover for 0, 4.5 or 7 h. Cells were then fixed, stained with rabbit polyclonal anti-DNA-PK CS pS2056, and counterstained with DAPI. At least 300 cells were scored for nuclei with > 5 DNA-PK CS foci. Error bars represent the standard error of the means from two independent experiments. ***, p

    Journal: PLoS ONE

    Article Title: Coordinate Nuclear Targeting of the FANCD2 and FANCI Proteins via a FANCD2 Nuclear Localization Signal

    doi: 10.1371/journal.pone.0081387

    Figure Lengend Snippet: The FANCD2 NLS mutants fail to correct the ICL sensitivity of FA-D2 patient cells. ( A ) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100, FANCD2-3N, or FANCD2-K561R were incubated in the absence or presence of 8 or 16 nM MMC for 24 h and the numbers of chromosome aberrations including gaps and breaks, dicentrics, and complex chromosome aberrations, including radial formations, were scored. Metaphase spreads were analyzed using a Zeiss AxioImager.A1 upright epifluorescent microscope with AxioVision LE 4.6 image acquisition software. At least 80 metaphases were scored per treatment. Error bars represent the standard error of the means. ( B ) FA-D2 cells stably expressing LacZ, FANCD2-WT and FANCD2-∆N57 were incubated in the absence (NT) or presence of 40 nM MMC for 18 h, and allowed to recover for 0, 4.5 or 7 h. Cells were then fixed, stained with rabbit polyclonal anti-DNA-PK CS pS2056, and counterstained with DAPI. At least 300 cells were scored for nuclei with > 5 DNA-PK CS foci. Error bars represent the standard error of the means from two independent experiments. ***, p

    Article Snippet: Nuclear foci were analyzed using a Zeiss AxioImager.A1 upright epifluorescent microscope with AxioVision LE 4.6 image acquisition software.

    Techniques: Stable Transfection, Expressing, Incubation, Microscopy, Software, Staining