human breast cancer cell line bt 474  (ATCC)


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    ATCC human breast cancer cell line bt 474
    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody"

    Article Title: Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody

    Journal: mAbs

    doi: 10.1080/19420862.2018.1486946

    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Figure Legend Snippet: ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.

    Techniques Used: Activity Assay

    human breast cancer cell lines bt 474  (ATCC)


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    ATCC human breast cancer cell lines bt 474
    Human Breast Cancer Cell Lines Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    human breast cancer cell line bt 474  (ATCC)


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    ATCC human breast cancer cell line bt 474
    Analysis of PPAB001 for simultaneous binding to CD47 and CD24 (A) Simultaneous binding of PPAB001 to recombinant CD47 and CD24 antigen by sandwich ELISA. (B) Flow cytometry of breast cancer <t>BT-474</t> cells stained with PPAB001, anti-CD24, CV1-hFc, or hIgG at 10 μg/mL. Binding kinetics of PPAB001, anti-CD24, or CV1-hFc to membrane-associated CD47 and CD24 on BT-474 cells were shown. Data are shown as means ± SDs (n = 3).
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    human breast cancer cell line bt 474 - by Bioz Stars, 2024-06
    86/100 stars

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    1) Product Images from "Dual blockade of CD47 and CD24 signaling using a novel bispecific antibody fusion protein enhances macrophage immunotherapy"

    Article Title: Dual blockade of CD47 and CD24 signaling using a novel bispecific antibody fusion protein enhances macrophage immunotherapy

    Journal: Molecular Therapy Oncolytics

    doi: 10.1016/j.omto.2023.100747

    Analysis of PPAB001 for simultaneous binding to CD47 and CD24 (A) Simultaneous binding of PPAB001 to recombinant CD47 and CD24 antigen by sandwich ELISA. (B) Flow cytometry of breast cancer BT-474 cells stained with PPAB001, anti-CD24, CV1-hFc, or hIgG at 10 μg/mL. Binding kinetics of PPAB001, anti-CD24, or CV1-hFc to membrane-associated CD47 and CD24 on BT-474 cells were shown. Data are shown as means ± SDs (n = 3).
    Figure Legend Snippet: Analysis of PPAB001 for simultaneous binding to CD47 and CD24 (A) Simultaneous binding of PPAB001 to recombinant CD47 and CD24 antigen by sandwich ELISA. (B) Flow cytometry of breast cancer BT-474 cells stained with PPAB001, anti-CD24, CV1-hFc, or hIgG at 10 μg/mL. Binding kinetics of PPAB001, anti-CD24, or CV1-hFc to membrane-associated CD47 and CD24 on BT-474 cells were shown. Data are shown as means ± SDs (n = 3).

    Techniques Used: Binding Assay, Recombinant, Sandwich ELISA, Flow Cytometry, Staining

    Treatment with PPAB001 promotes phagocytosis of human cancer cells (A) Representative phagocytosis images of macrophages engulfing BT-474, BT-474R, or SK-OV-3 cells with PPAB001, anti-CD24, CV1-hFc, or hIgG. Macrophages were stained red (CMTPX); cancer cells were labeled with green (CFSE). The white arrows indicate macrophages that engulfed cancer cells. (B) The extent of macrophage phagocytosis with PPAB001, anti-CD24, CV1-hFc, or hIgG was quantified for BT-474, BT-474R, or SK-OV-3 cells. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.
    Figure Legend Snippet: Treatment with PPAB001 promotes phagocytosis of human cancer cells (A) Representative phagocytosis images of macrophages engulfing BT-474, BT-474R, or SK-OV-3 cells with PPAB001, anti-CD24, CV1-hFc, or hIgG. Macrophages were stained red (CMTPX); cancer cells were labeled with green (CFSE). The white arrows indicate macrophages that engulfed cancer cells. (B) The extent of macrophage phagocytosis with PPAB001, anti-CD24, CV1-hFc, or hIgG was quantified for BT-474, BT-474R, or SK-OV-3 cells. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

    Techniques Used: Staining, Labeling

    CD47/CD24 dual blockage potentiates the macrophage phagocytosis (A) Representative flow cytometry plots depicting the phagocytosis of BT-474 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (B) Representative flow cytometry plots depicting the phagocytosis of BT-474R cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (C) Representative flow cytometry plots depicting the phagocytosis of SK-OV-3 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.
    Figure Legend Snippet: CD47/CD24 dual blockage potentiates the macrophage phagocytosis (A) Representative flow cytometry plots depicting the phagocytosis of BT-474 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (B) Representative flow cytometry plots depicting the phagocytosis of BT-474R cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (C) Representative flow cytometry plots depicting the phagocytosis of SK-OV-3 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

    Techniques Used: Flow Cytometry

    human breast cancer bt 474 cell lines  (ATCC)


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    ATCC human breast cancer bt 474 cell lines
    Human Breast Cancer Bt 474 Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human breast cancer cell line bt 474  (ATCC)


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    ATCC human breast cancer cell line bt 474
    (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing <t>BT-474</t> cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Lysine-Directed Site-Selective Bioconjugation for the Creation of Radioimmunoconjugates"

    Article Title: Lysine-Directed Site-Selective Bioconjugation for the Creation of Radioimmunoconjugates

    Journal: Bioconjugate chemistry

    doi: 10.1021/acs.bioconjchem.2c00354

    (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing BT-474 cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.
    Figure Legend Snippet: (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing BT-474 cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.

    Techniques Used: Fluorescence, FACS, Expressing, Labeling, Magnetic Beads, Two Tailed Test, Software

    (A) Representative PET scans collected 24, 48, 72, 96, 120, and 144 h after the intravenous administration of [89Z]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, or [89Zr]Zr-malDFO-pertuzumab [3.7 −3.9 MBq (100-105 μCi), 20-21 μg, in 100 μL of PBS] to athymic nude mice bearing subcutaneous HER2-expressing BT-474 human breast cancer xenografts (n = 5). The images on the left are coronal slices, and those on the right are maximum intensity projections (MIPs); (B) biodistribution data from the mice used for PET imaging collected after the terminal imaging timepoint at 144 h (n = 5).
    Figure Legend Snippet: (A) Representative PET scans collected 24, 48, 72, 96, 120, and 144 h after the intravenous administration of [89Z]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, or [89Zr]Zr-malDFO-pertuzumab [3.7 −3.9 MBq (100-105 μCi), 20-21 μg, in 100 μL of PBS] to athymic nude mice bearing subcutaneous HER2-expressing BT-474 human breast cancer xenografts (n = 5). The images on the left are coronal slices, and those on the right are maximum intensity projections (MIPs); (B) biodistribution data from the mice used for PET imaging collected after the terminal imaging timepoint at 144 h (n = 5).

    Techniques Used: Expressing, Imaging

    human breast cancer cell line bt 474  (ATCC)


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    ATCC human breast cancer cell line bt 474
    (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing <t>BT-474</t> cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human breast cancer cell line bt 474/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human breast cancer cell line bt 474 - by Bioz Stars, 2024-06
    86/100 stars

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    1) Product Images from "Lysine-Directed Site-Selective Bioconjugation for the Creation of Radioimmunoconjugates"

    Article Title: Lysine-Directed Site-Selective Bioconjugation for the Creation of Radioimmunoconjugates

    Journal: Bioconjugate chemistry

    doi: 10.1021/acs.bioconjchem.2c00354

    (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing BT-474 cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.
    Figure Legend Snippet: (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing BT-474 cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.

    Techniques Used: Fluorescence, FACS, Expressing, Labeling, Magnetic Beads, Two Tailed Test, Software

    (A) Representative PET scans collected 24, 48, 72, 96, 120, and 144 h after the intravenous administration of [89Z]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, or [89Zr]Zr-malDFO-pertuzumab [3.7 −3.9 MBq (100-105 μCi), 20-21 μg, in 100 μL of PBS] to athymic nude mice bearing subcutaneous HER2-expressing BT-474 human breast cancer xenografts (n = 5). The images on the left are coronal slices, and those on the right are maximum intensity projections (MIPs); (B) biodistribution data from the mice used for PET imaging collected after the terminal imaging timepoint at 144 h (n = 5).
    Figure Legend Snippet: (A) Representative PET scans collected 24, 48, 72, 96, 120, and 144 h after the intravenous administration of [89Z]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, or [89Zr]Zr-malDFO-pertuzumab [3.7 −3.9 MBq (100-105 μCi), 20-21 μg, in 100 μL of PBS] to athymic nude mice bearing subcutaneous HER2-expressing BT-474 human breast cancer xenografts (n = 5). The images on the left are coronal slices, and those on the right are maximum intensity projections (MIPs); (B) biodistribution data from the mice used for PET imaging collected after the terminal imaging timepoint at 144 h (n = 5).

    Techniques Used: Expressing, Imaging

    human breast cancer cell line bt 474  (ATCC)


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    ATCC human breast cancer cell line bt 474
    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human breast cancer cell line bt 474/product/ATCC
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human breast cancer cell line bt 474 - by Bioz Stars, 2024-06
    93/100 stars

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    1) Product Images from "Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody"

    Article Title: Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody

    Journal: mAbs

    doi: 10.1080/19420862.2018.1486946

    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Figure Legend Snippet: ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.

    Techniques Used: Activity Assay

    human breast cancer cell lines bt 474  (ATCC)


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    ATCC human breast cancer cell lines bt 474
    Expression of HER2 in breast cancer cells, analyzed by flow cytometry. Notes: Representative fluorescence-activated cell sorting analysis of ( A ) MDA-MB-361 and ( B ) <t>BT-474</t> cells tested by ALDEFLUOR assay. In the right image (ALDH+ DEAB), cells incubated with ALDH substrate (BAAA) and the specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells (R1) and to define the ALDEFLUOR-positive region (R2). ( C ) Percentage of HER2 positively stained cells in breast cancer cells. ( D ) HER2 mean fluorescence intensity in breast cancer cells. Data are expressed as mean ± SD (n=3). ** P <0.01. Abbreviations: ALDH, aldehyde dehydrogenase; BAAA, BODIPY-aminoacetaldehyde; DEAB, diethylaminobenzaldehyde.
    Human Breast Cancer Cell Lines Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human breast cancer cell lines bt 474/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human breast cancer cell lines bt 474 - by Bioz Stars, 2024-06
    86/100 stars

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    1) Product Images from "Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells"

    Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S144184

    Expression of HER2 in breast cancer cells, analyzed by flow cytometry. Notes: Representative fluorescence-activated cell sorting analysis of ( A ) MDA-MB-361 and ( B ) BT-474 cells tested by ALDEFLUOR assay. In the right image (ALDH+ DEAB), cells incubated with ALDH substrate (BAAA) and the specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells (R1) and to define the ALDEFLUOR-positive region (R2). ( C ) Percentage of HER2 positively stained cells in breast cancer cells. ( D ) HER2 mean fluorescence intensity in breast cancer cells. Data are expressed as mean ± SD (n=3). ** P <0.01. Abbreviations: ALDH, aldehyde dehydrogenase; BAAA, BODIPY-aminoacetaldehyde; DEAB, diethylaminobenzaldehyde.
    Figure Legend Snippet: Expression of HER2 in breast cancer cells, analyzed by flow cytometry. Notes: Representative fluorescence-activated cell sorting analysis of ( A ) MDA-MB-361 and ( B ) BT-474 cells tested by ALDEFLUOR assay. In the right image (ALDH+ DEAB), cells incubated with ALDH substrate (BAAA) and the specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells (R1) and to define the ALDEFLUOR-positive region (R2). ( C ) Percentage of HER2 positively stained cells in breast cancer cells. ( D ) HER2 mean fluorescence intensity in breast cancer cells. Data are expressed as mean ± SD (n=3). ** P <0.01. Abbreviations: ALDH, aldehyde dehydrogenase; BAAA, BODIPY-aminoacetaldehyde; DEAB, diethylaminobenzaldehyde.

    Techniques Used: Expressing, Flow Cytometry, Fluorescence, FACS, Incubation, Staining

    Cell proliferation assay. Notes: ALDH+ and ALDH− breast cancer cells were seeded in 96-well plates with a density of 1×10 4 cells per well overnight. The cells were incubated with the nanoparticles or salinomycin for 72 h, and cell viability was evaluated using the CCK-8 assay. ( A ) MDA-MB-361 ALDH+, ( B ) MDA-MB-361 ALDH−, ( C ) BT-474 ALDH+, and ( D ) BT-474 ALDH−. Data are expressed as mean ± SD (n=3). Abbreviations: ALDH, aldehyde dehydrogenase; CCK-8, Cell Counting Kit-8; Sali-NP, salinomycin-loaded polymer-lipid nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.
    Figure Legend Snippet: Cell proliferation assay. Notes: ALDH+ and ALDH− breast cancer cells were seeded in 96-well plates with a density of 1×10 4 cells per well overnight. The cells were incubated with the nanoparticles or salinomycin for 72 h, and cell viability was evaluated using the CCK-8 assay. ( A ) MDA-MB-361 ALDH+, ( B ) MDA-MB-361 ALDH−, ( C ) BT-474 ALDH+, and ( D ) BT-474 ALDH−. Data are expressed as mean ± SD (n=3). Abbreviations: ALDH, aldehyde dehydrogenase; CCK-8, Cell Counting Kit-8; Sali-NP, salinomycin-loaded polymer-lipid nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Techniques Used: Proliferation Assay, Incubation, CCK-8 Assay, Cell Counting

    The IC 50 values of salinomycin and nanoparticles in breast cancer cells
    Figure Legend Snippet: The IC 50 values of salinomycin and nanoparticles in breast cancer cells

    Techniques Used:

    Effect of treatments on the percentage of CSCs. Notes: Effect of the treatments on the proportion of CSCs in breast cancer cells, as reflected by the tumorsphere formation rate ( A and B ) and the proportion of ALDH+ cells ( E and F ). Representative images of tumorspheres formed by ( C ) MDA-MB-361 cells and ( D ) BT-474 cells are shown. The rate of tumorsphere formation is defined as the number of tumorspheres formed in 7 days in the treatment group divided by the number of tumorspheres formed in 7 days in the untreated group; the rate of tumorsphere formation in the untreated group is used as a control and defined as 100%. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=6). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSCs, cancer stem cells; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.
    Figure Legend Snippet: Effect of treatments on the percentage of CSCs. Notes: Effect of the treatments on the proportion of CSCs in breast cancer cells, as reflected by the tumorsphere formation rate ( A and B ) and the proportion of ALDH+ cells ( E and F ). Representative images of tumorspheres formed by ( C ) MDA-MB-361 cells and ( D ) BT-474 cells are shown. The rate of tumorsphere formation is defined as the number of tumorspheres formed in 7 days in the treatment group divided by the number of tumorspheres formed in 7 days in the untreated group; the rate of tumorsphere formation in the untreated group is used as a control and defined as 100%. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=6). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSCs, cancer stem cells; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Techniques Used:

    In vivo antitumor assay in mice bearing subcutaneous BT-474 tumors. Notes: BT-474 tumors reached ~50 mm 3 in size by day 10. From day 10, mice were injected with nanoparticles (7.5 mg salinomycin/kg, i.v.) through the tail vein, and salinomycin (7.5 mg salinomycin/kg) dissolved in ethanol was administered by intraperitoneal injection. Therapy was given nine times on alternate days (indicated by arrows), and tumor volume was calculated. ( A ) Tumor growth curve. ( B ) Excised tumors. ( C ) The excised tumors were weighed at the end point. On day 28, the effect of the drug treatments on the CSC proportion in BT-474 tumors in vivo was evaluated by ( D ) the rate of tumorsphere formation and ( E ) the proportion of ALDH+ cells from the excised tumors. ( F ) Representative images of tumorspheres from ( D ) are shown. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=8). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSC, cancer stem cell; i.v., intravenous; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.
    Figure Legend Snippet: In vivo antitumor assay in mice bearing subcutaneous BT-474 tumors. Notes: BT-474 tumors reached ~50 mm 3 in size by day 10. From day 10, mice were injected with nanoparticles (7.5 mg salinomycin/kg, i.v.) through the tail vein, and salinomycin (7.5 mg salinomycin/kg) dissolved in ethanol was administered by intraperitoneal injection. Therapy was given nine times on alternate days (indicated by arrows), and tumor volume was calculated. ( A ) Tumor growth curve. ( B ) Excised tumors. ( C ) The excised tumors were weighed at the end point. On day 28, the effect of the drug treatments on the CSC proportion in BT-474 tumors in vivo was evaluated by ( D ) the rate of tumorsphere formation and ( E ) the proportion of ALDH+ cells from the excised tumors. ( F ) Representative images of tumorspheres from ( D ) are shown. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=8). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSC, cancer stem cell; i.v., intravenous; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Techniques Used: In Vivo, Injection

    human bt474 breast cancer cell line  (ATCC)


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    Structured Review

    ATCC human bt474 breast cancer cell line
    Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the <t>BT474</t> breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)
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    1) Product Images from "Phosphorylation of Ser 78 of Hsp27 correlated with HER-2/ neu status and lymph node positivity in breast cancer"

    Article Title: Phosphorylation of Ser 78 of Hsp27 correlated with HER-2/ neu status and lymph node positivity in breast cancer

    Journal: Molecular Cancer

    doi: 10.1186/1476-4598-6-52

    Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the BT474 breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)
    Figure Legend Snippet: Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the BT474 breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)

    Techniques Used: SDS Page, Cell Culture

    human breast cancer cell lines bt 474  (ATCC)


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    ATCC human breast cancer cell lines bt 474
    Expression levels of Bcl-2 and Bcl-xL proteins in MDA-MB-231, MDA-MD-453, <t>BT-474,</t> and ZR-75-1 cells. (a) Western blot analysis of Bcl-2 and Bcl-xL expression. (b) Quantification of Bcl-2 and Bcl-xL expression by densitometric analysis. The relative expression of Bcl-2 and Bcl-xL in MDA-MB-453 cells was compared with the expression in MDA-MB-231, BT-474, and ZR-75-1 cells. Results are from two representative, independent experiments.
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    1) Product Images from "Targeted therapy against Bcl-2-related proteins in breast cancer cells"

    Article Title: Targeted therapy against Bcl-2-related proteins in breast cancer cells

    Journal: Breast Cancer Research

    doi: 10.1186/bcr1323

    Expression levels of Bcl-2 and Bcl-xL proteins in MDA-MB-231, MDA-MD-453, BT-474, and ZR-75-1 cells. (a) Western blot analysis of Bcl-2 and Bcl-xL expression. (b) Quantification of Bcl-2 and Bcl-xL expression by densitometric analysis. The relative expression of Bcl-2 and Bcl-xL in MDA-MB-453 cells was compared with the expression in MDA-MB-231, BT-474, and ZR-75-1 cells. Results are from two representative, independent experiments.
    Figure Legend Snippet: Expression levels of Bcl-2 and Bcl-xL proteins in MDA-MB-231, MDA-MD-453, BT-474, and ZR-75-1 cells. (a) Western blot analysis of Bcl-2 and Bcl-xL expression. (b) Quantification of Bcl-2 and Bcl-xL expression by densitometric analysis. The relative expression of Bcl-2 and Bcl-xL in MDA-MB-453 cells was compared with the expression in MDA-MB-231, BT-474, and ZR-75-1 cells. Results are from two representative, independent experiments.

    Techniques Used: Expressing, Western Blot

    Sequence-specific downregulation and cytotoxic effects of antisense Bcl-2 oligodeoxynucleotides on BT-474 and ZR-75-1 cells. (a) Specific inhibition of Bcl-2 protein expression by treatment with antisense (AS) Bcl-2 oligodeoxynucleotides (ODNs). Cells were treated with 10 μg/ml Lipofectamine alone (control) or 1.0 μM AS, mismatch control (MM), or random control (RC) ODNs for 24 hours. Cells were then cultured in standard medium, total protein was extracted, and Bcl-2 and β-actin protein levels were analyzed by Western blotting. (b) Quantification of Bcl-2 expression by densitometric analysis. The expression of Bcl-2 in cells treated with control, AS Bcl-2 , RC Bcl-2 , and MM Bcl-2 ODNs was normalized with β-actin, and the relative values are presented. Error bars indicate SD. The data presented are from three independent experiments. (c) Effects of AS Bcl-2 ODNs on the proliferation of BT-474 and ZR-75-1 breast cancer cells in vitro . Cells were treated with various concentrations of AS Bcl-2 ODNs in 24-well dishes. Four days after treatment, cells were stained with trypan blue and counted. Error bars indicate SD. The data presented are from three independent experiments.
    Figure Legend Snippet: Sequence-specific downregulation and cytotoxic effects of antisense Bcl-2 oligodeoxynucleotides on BT-474 and ZR-75-1 cells. (a) Specific inhibition of Bcl-2 protein expression by treatment with antisense (AS) Bcl-2 oligodeoxynucleotides (ODNs). Cells were treated with 10 μg/ml Lipofectamine alone (control) or 1.0 μM AS, mismatch control (MM), or random control (RC) ODNs for 24 hours. Cells were then cultured in standard medium, total protein was extracted, and Bcl-2 and β-actin protein levels were analyzed by Western blotting. (b) Quantification of Bcl-2 expression by densitometric analysis. The expression of Bcl-2 in cells treated with control, AS Bcl-2 , RC Bcl-2 , and MM Bcl-2 ODNs was normalized with β-actin, and the relative values are presented. Error bars indicate SD. The data presented are from three independent experiments. (c) Effects of AS Bcl-2 ODNs on the proliferation of BT-474 and ZR-75-1 breast cancer cells in vitro . Cells were treated with various concentrations of AS Bcl-2 ODNs in 24-well dishes. Four days after treatment, cells were stained with trypan blue and counted. Error bars indicate SD. The data presented are from three independent experiments.

    Techniques Used: Sequencing, Inhibition, Expressing, Cell Culture, Western Blot, In Vitro, Staining

    Effect of antisense Bcl-2 oligodeoxynucleotides on chemosensitivity in  BT-474,  ZR-75-1, and MDA-MB-231 breast cancer cells
    Figure Legend Snippet: Effect of antisense Bcl-2 oligodeoxynucleotides on chemosensitivity in BT-474, ZR-75-1, and MDA-MB-231 breast cancer cells

    Techniques Used:

    Sequence-specific downregulation and cytotoxic effects of antisense Bcl-xL oligodeoxynucleotides on MDA-MB-231 and BT-474 cells. (a) Specific inhibition of Bcl-xL protein expression by treatment with antisense (AS) Bcl-xL oligodeoxynucleotides (ODNs). Cells were treated with 10 μg/ml Lipofectamine alone (control) or 1.0 μM AS, mismatch control (MM), or random control (RC) ODNs for 24 hours. Cells were then cultured in standard medium, total protein was extracted, and Bcl-xL and β-actin protein levels were analyzed by Western blotting. (b) Quantification of Bcl-xL protein expression by densitometric analysis. The Bcl-xL protein expression was normalized with β-actin, and the relative values are presented. Error bars indicate SD. The data presented are from three independent experiments. (c) Effects of AS Bcl-xL ODNs on the proliferation of MDA-MB-231 and BT-474 breast cancer cells in vitro . Cells were treated with various concentrations of AS Bcl-xL ODNs in 24-well dishes. Four days after treatment, cells were stained with trypan blue and counted. Error bars indicate SD. The data presented are from three independent experiments.
    Figure Legend Snippet: Sequence-specific downregulation and cytotoxic effects of antisense Bcl-xL oligodeoxynucleotides on MDA-MB-231 and BT-474 cells. (a) Specific inhibition of Bcl-xL protein expression by treatment with antisense (AS) Bcl-xL oligodeoxynucleotides (ODNs). Cells were treated with 10 μg/ml Lipofectamine alone (control) or 1.0 μM AS, mismatch control (MM), or random control (RC) ODNs for 24 hours. Cells were then cultured in standard medium, total protein was extracted, and Bcl-xL and β-actin protein levels were analyzed by Western blotting. (b) Quantification of Bcl-xL protein expression by densitometric analysis. The Bcl-xL protein expression was normalized with β-actin, and the relative values are presented. Error bars indicate SD. The data presented are from three independent experiments. (c) Effects of AS Bcl-xL ODNs on the proliferation of MDA-MB-231 and BT-474 breast cancer cells in vitro . Cells were treated with various concentrations of AS Bcl-xL ODNs in 24-well dishes. Four days after treatment, cells were stained with trypan blue and counted. Error bars indicate SD. The data presented are from three independent experiments.

    Techniques Used: Sequencing, Inhibition, Expressing, Cell Culture, Western Blot, In Vitro, Staining

    Effect of antisense Bcl-xL oligodeoxynucleotides on chemosensitivity in  BT-474,  ZR-75-1, and MDA-MB-231 breast cancer cells
    Figure Legend Snippet: Effect of antisense Bcl-xL oligodeoxynucleotides on chemosensitivity in BT-474, ZR-75-1, and MDA-MB-231 breast cancer cells

    Techniques Used:

    Effects of treatment with antisense Bcl-2 and mitomycin C, doxorubicin, paclitaxel, or docetaxel on BT-474 cells. (a) Expression levels of Bcl-2 and Bcl-xL protein in BT-474 cells transplanted into athymic mice after treatment with antisense (AS) Bcl-2 oligodeoxynucleotides (ODNs) were measured by Western blot analysis at the indicated time points. (b) Enhancement of the antitumor effects of anticancer drugs by AS Bcl-2 ODNs in BT-474 tumor xenografts. Each point represents the mean tumor volume of the eight mice in each group. Error bars indicate SD. *, P < 0.05, analysis of variance with Fisher's least significant difference test. The data presented are from two independent experiments. MMC, mitomycin C; DOX, doxorubicin; TXL, paclitaxel; TXT, docetaxel.
    Figure Legend Snippet: Effects of treatment with antisense Bcl-2 and mitomycin C, doxorubicin, paclitaxel, or docetaxel on BT-474 cells. (a) Expression levels of Bcl-2 and Bcl-xL protein in BT-474 cells transplanted into athymic mice after treatment with antisense (AS) Bcl-2 oligodeoxynucleotides (ODNs) were measured by Western blot analysis at the indicated time points. (b) Enhancement of the antitumor effects of anticancer drugs by AS Bcl-2 ODNs in BT-474 tumor xenografts. Each point represents the mean tumor volume of the eight mice in each group. Error bars indicate SD. *, P < 0.05, analysis of variance with Fisher's least significant difference test. The data presented are from two independent experiments. MMC, mitomycin C; DOX, doxorubicin; TXL, paclitaxel; TXT, docetaxel.

    Techniques Used: Expressing, Western Blot

    Effect of synthetic CpG antisense Bcl-2 on BT-474 cells in comparison with antisense Bcl-2 . Each point represents the mean tumor volume of the four mice in each group. Error bars indicate SD. The data presented are from two independent experiments. RC, random control; TXT, docetaxel.
    Figure Legend Snippet: Effect of synthetic CpG antisense Bcl-2 on BT-474 cells in comparison with antisense Bcl-2 . Each point represents the mean tumor volume of the four mice in each group. Error bars indicate SD. The data presented are from two independent experiments. RC, random control; TXT, docetaxel.

    Techniques Used:

    human breast cancer cell lines bt 474  (ATCC)


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    ATCC human breast cancer cell lines bt 474
    Plot of the (total/bound) activity versus (1/[normalized cell concentration]), used to calculate the immunoreactivity fraction of 89 Zr-DFO-trastuzumab in <t>BT-474</t> (HER2/ neu positive) cells by extrapolation to infinite antigen excess (1/ y -intercept).
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    1) Product Images from "Measuring the Pharmacodynamic Effects of a Novel Hsp90 Inhibitor on HER2/ neu Expression in Mice Using 89 Zr-DFO-Trastuzumab"

    Article Title: Measuring the Pharmacodynamic Effects of a Novel Hsp90 Inhibitor on HER2/ neu Expression in Mice Using 89 Zr-DFO-Trastuzumab

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0008859

    Plot of the (total/bound) activity versus (1/[normalized cell concentration]), used to calculate the immunoreactivity fraction of 89 Zr-DFO-trastuzumab in BT-474 (HER2/ neu positive) cells by extrapolation to infinite antigen excess (1/ y -intercept).
    Figure Legend Snippet: Plot of the (total/bound) activity versus (1/[normalized cell concentration]), used to calculate the immunoreactivity fraction of 89 Zr-DFO-trastuzumab in BT-474 (HER2/ neu positive) cells by extrapolation to infinite antigen excess (1/ y -intercept).

    Techniques Used: Activity Assay, Concentration Assay

    Biodistribution data of 89 Zr-DFO-trastuzumab versus time/h, administered by i.v. tail-vein injection to female, athymic nu / nu mice bearing s.c.  BT-474  tumors (90–150 mm 3 ). <xref ref-type= a,b " title="... female, athymic nu / nu mice bearing s.c. BT-474 tumors (90–150 mm 3 ).
    Figure Legend Snippet: Biodistribution data of 89 Zr-DFO-trastuzumab versus time/h, administered by i.v. tail-vein injection to female, athymic nu / nu mice bearing s.c. BT-474 tumors (90–150 mm 3 ). a,b

    Techniques Used: Injection

    Bar charts showing selected tissue biodistribution data (%ID/g) for (A) uptake of high and low specific-activity formulations of 89 Zr-DFO-trastuzumab in BT-474 tumor-bearing mice, and (B) 89 Zr-DFO-trastuzumab uptake in control (vehicle-treated) and PU-H71 treated animals at 12, 24, 48 and 72 h post-i.v. administration of 89 Zr-DFO-trastuzumab (0.55–0.74 MBq, 5–7 µg of mAb, in 200 µL 0.9% sterile saline).
    Figure Legend Snippet: Bar charts showing selected tissue biodistribution data (%ID/g) for (A) uptake of high and low specific-activity formulations of 89 Zr-DFO-trastuzumab in BT-474 tumor-bearing mice, and (B) 89 Zr-DFO-trastuzumab uptake in control (vehicle-treated) and PU-H71 treated animals at 12, 24, 48 and 72 h post-i.v. administration of 89 Zr-DFO-trastuzumab (0.55–0.74 MBq, 5–7 µg of mAb, in 200 µL 0.9% sterile saline).

    Techniques Used: Activity Assay

    Pharmacodynamic studies on protein expression levels in BT-474 tumor tissue samples obtained at 12, 24, 48, 72 and 96 h after PU-H71 treatment.
    Figure Legend Snippet: Pharmacodynamic studies on protein expression levels in BT-474 tumor tissue samples obtained at 12, 24, 48, 72 and 96 h after PU-H71 treatment.

    Techniques Used: Expressing

    Time-activity curves derived by region-of-interest analysis of the immunoPET images showing the mean %ID/g tissue uptake versus time/h, for control and PU-H71-treated mice bearing both BT-474 and MDA-MB-468 tumors.
    Figure Legend Snippet: Time-activity curves derived by region-of-interest analysis of the immunoPET images showing the mean %ID/g tissue uptake versus time/h, for control and PU-H71-treated mice bearing both BT-474 and MDA-MB-468 tumors.

    Techniques Used: Activity Assay, Derivative Assay

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    ATCC human breast cancer cell line bt 474
    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human breast cancer cell lines bt 474
    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Human Breast Cancer Cell Lines Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human breast cancer bt 474 cell lines
    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Human Breast Cancer Bt 474 Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human bt474 breast cancer cell line
    Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the <t>BT474</t> breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)
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    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.

    Journal: mAbs

    Article Title: Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody

    doi: 10.1080/19420862.2018.1486946

    Figure Lengend Snippet: ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.

    Article Snippet: Human breast cancer cell line BT-474 (HTB-20), MDA-MB-175VII (HTB-25), SK-BR-3(HTB-30) and HCC1419(CRL-2326) are HER2-positive cell lines, which were obtained from the ATCC, and HCC1419 is trastuzumab-resistant human breast cancer cell line.

    Techniques: Activity Assay

    Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the BT474 breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)

    Journal: Molecular Cancer

    Article Title: Phosphorylation of Ser 78 of Hsp27 correlated with HER-2/ neu status and lymph node positivity in breast cancer

    doi: 10.1186/1476-4598-6-52

    Figure Lengend Snippet: Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the BT474 breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)

    Article Snippet: The human BT474 breast cancer cell line was obtained from American Type Culture Collection, and maintained in modified Dulbecco's medium (HybriCare) supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified atmosphere of 5% CO 2 in an incubator.

    Techniques: SDS Page, Cell Culture