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  • 99
    Thermo Fisher foetal bovine serum fbs
    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered <t>DMEM</t> (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% <t>FBS.</t> The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.
    Foetal Bovine Serum Fbs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3259 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore fbv solution
    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered <t>DMEM</t> (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% <t>FBS.</t> The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.
    Fbv Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Gilead Sciences fbv
    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered <t>DMEM</t> (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% <t>FBS.</t> The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.
    Fbv, supplied by Gilead Sciences, 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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    86
    The Jackson Laboratory wild type fbv mice
    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered <t>DMEM</t> (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% <t>FBS.</t> The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.
    Wild Type Fbv Mice, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    The Jackson Laboratory b6 fbv tg
    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered <t>DMEM</t> (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% <t>FBS.</t> The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.
    B6 Fbv Tg, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    GE Healthcare blood velocity fbv
    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered <t>DMEM</t> (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% <t>FBS.</t> The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.
    Blood Velocity Fbv, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Olympus olympus fbv 1000
    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered <t>DMEM</t> (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% <t>FBS.</t> The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.
    Olympus Fbv 1000, supplied by Olympus, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Charles River Laboratories female fbv mice
    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered <t>DMEM</t> (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% <t>FBS.</t> The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.
    Female Fbv Mice, supplied by Charles River Laboratories, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    GE Healthcare fbs fbs
    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered <t>DMEM</t> (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% <t>FBS.</t> The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.
    Fbs Fbs, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher fbv n control mice
    qRT-PCR validation of the GEP microarray data ( blue ) in Sca1+ cells purified from BM or spleen of another set of <t>HR-MDS</t> mice ( two different hues of orange ), and CD34+ cells purified from MDS patients ( green ) compared to <t>FBV/N</t> or CD34 BM cells, respectively. Data expressed as fold change of expression of several genes of three pathways shown (angpt1 (signal transduction); cox4i1 and ndufv2 (oxidative metabolism); sdc1 (DNA processing)). qRT-PCR fold changes shown here are represented as the average of three experiments
    Fbv N Control Mice, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher fbs fbs
    Overexpression of 70z-Cbl promotes the loss of cell–cell contacts. (A) Parental <t>MDCK</t> cells, unstimulated or stimulated with 5 U/ml HGF for 24 h, as well as stable cell lines expressing c-Cbl (clone 8) and 70z-Cbl (clone 20), were grown on glass coverslips in DMEM containing 10% <t>FBS.</t> Cells were treated for 10 min with CSK buffer, fixed in 3.7% formaldehyde, and then labeled with anti-E-cadherin antibody followed by Cy3-conjugated anti-mouse antiserum (a, c, e, and g). Matching phase-contrast images are shown (b, d, f, and h). Photographs were taken at a magnification of ×63. (B) Whole cell lysates were separated on an 8% SDS-PAGE gel and immunoblotted with anti-E-cadherin antibody.
    Fbs Fbs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11657 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Biological Industries Inc fbs cd fbs
    Overexpression of 70z-Cbl promotes the loss of cell–cell contacts. (A) Parental <t>MDCK</t> cells, unstimulated or stimulated with 5 U/ml HGF for 24 h, as well as stable cell lines expressing c-Cbl (clone 8) and 70z-Cbl (clone 20), were grown on glass coverslips in DMEM containing 10% <t>FBS.</t> Cells were treated for 10 min with CSK buffer, fixed in 3.7% formaldehyde, and then labeled with anti-E-cadherin antibody followed by Cy3-conjugated anti-mouse antiserum (a, c, e, and g). Matching phase-contrast images are shown (b, d, f, and h). Photographs were taken at a magnification of ×63. (B) Whole cell lysates were separated on an 8% SDS-PAGE gel and immunoblotted with anti-E-cadherin antibody.
    Fbs Cd Fbs, supplied by Biological Industries Inc, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    The Jackson Laboratory wild type controls fbv nj
    Overexpression of 70z-Cbl promotes the loss of cell–cell contacts. (A) Parental <t>MDCK</t> cells, unstimulated or stimulated with 5 U/ml HGF for 24 h, as well as stable cell lines expressing c-Cbl (clone 8) and 70z-Cbl (clone 20), were grown on glass coverslips in DMEM containing 10% <t>FBS.</t> Cells were treated for 10 min with CSK buffer, fixed in 3.7% formaldehyde, and then labeled with anti-E-cadherin antibody followed by Cy3-conjugated anti-mouse antiserum (a, c, e, and g). Matching phase-contrast images are shown (b, d, f, and h). Photographs were taken at a magnification of ×63. (B) Whole cell lysates were separated on an 8% SDS-PAGE gel and immunoblotted with anti-E-cadherin antibody.
    Wild Type Controls Fbv Nj, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 85 stars, based on 1 article reviews
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    92
    Biochrom fbs cs fbs
    Overexpression of 70z-Cbl promotes the loss of cell–cell contacts. (A) Parental <t>MDCK</t> cells, unstimulated or stimulated with 5 U/ml HGF for 24 h, as well as stable cell lines expressing c-Cbl (clone 8) and 70z-Cbl (clone 20), were grown on glass coverslips in DMEM containing 10% <t>FBS.</t> Cells were treated for 10 min with CSK buffer, fixed in 3.7% formaldehyde, and then labeled with anti-E-cadherin antibody followed by Cy3-conjugated anti-mouse antiserum (a, c, e, and g). Matching phase-contrast images are shown (b, d, f, and h). Photographs were taken at a magnification of ×63. (B) Whole cell lysates were separated on an 8% SDS-PAGE gel and immunoblotted with anti-E-cadherin antibody.
    Fbs Cs Fbs, supplied by Biochrom, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bio-Rad fetal bovine serum
    Overexpression of 70z-Cbl promotes the loss of cell–cell contacts. (A) Parental <t>MDCK</t> cells, unstimulated or stimulated with 5 U/ml HGF for 24 h, as well as stable cell lines expressing c-Cbl (clone 8) and 70z-Cbl (clone 20), were grown on glass coverslips in DMEM containing 10% <t>FBS.</t> Cells were treated for 10 min with CSK buffer, fixed in 3.7% formaldehyde, and then labeled with anti-E-cadherin antibody followed by Cy3-conjugated anti-mouse antiserum (a, c, e, and g). Matching phase-contrast images are shown (b, d, f, and h). Photographs were taken at a magnification of ×63. (B) Whole cell lysates were separated on an 8% SDS-PAGE gel and immunoblotted with anti-E-cadherin antibody.
    Fetal Bovine Serum, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 235 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    fbs  (Lonza)
    97
    Lonza fbs
    Tm7sf2 gene presides over an anti inflammatory loop. (a) Nuclear translocation of Nrf2 and NF-κB in WT and KO <t>MEFs.</t> MEFs, grown in DMEM plus 5%LPDS, were treated with 1 µM thapsigargin. At the indicated times, cells were collected and nuclear extracts subjected to Western blotting analyses with the indicated antibodies. Lamin B was used as loading control; (b) TNFα and HO-1 gene expression in WT and KO MEFs. MEFs, grown in DMEM plus 5%LPDS, were treated with 1 µM thapsigargin for 6 hr and subjected to real time PCR analysis. (c) TNFα expression in KO MEFs grown in <t>10%FBS</t> and treated with 1 µM thapsigargin for 6 hr and subjected to real time PCR analysis. Expression of each gene was normalized to GAPDH and reported as 2 −ΔΔCt . Relative mRNA level of WT untreated cells was assumed as 1. Results are given as mean ± s.d., (n = 7). *p
    Fbs, supplied by Lonza, used in various techniques. Bioz Stars score: 97/100, based on 1981 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Mediatech fbs rpmi
    Tm7sf2 gene presides over an anti inflammatory loop. (a) Nuclear translocation of Nrf2 and NF-κB in WT and KO <t>MEFs.</t> MEFs, grown in DMEM plus 5%LPDS, were treated with 1 µM thapsigargin. At the indicated times, cells were collected and nuclear extracts subjected to Western blotting analyses with the indicated antibodies. Lamin B was used as loading control; (b) TNFα and HO-1 gene expression in WT and KO MEFs. MEFs, grown in DMEM plus 5%LPDS, were treated with 1 µM thapsigargin for 6 hr and subjected to real time PCR analysis. (c) TNFα expression in KO MEFs grown in <t>10%FBS</t> and treated with 1 µM thapsigargin for 6 hr and subjected to real time PCR analysis. Expression of each gene was normalized to GAPDH and reported as 2 −ΔΔCt . Relative mRNA level of WT untreated cells was assumed as 1. Results are given as mean ± s.d., (n = 7). *p
    Fbs Rpmi, supplied by Mediatech, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Becton Dickinson buffer fbs
    Tm7sf2 gene presides over an anti inflammatory loop. (a) Nuclear translocation of Nrf2 and NF-κB in WT and KO <t>MEFs.</t> MEFs, grown in DMEM plus 5%LPDS, were treated with 1 µM thapsigargin. At the indicated times, cells were collected and nuclear extracts subjected to Western blotting analyses with the indicated antibodies. Lamin B was used as loading control; (b) TNFα and HO-1 gene expression in WT and KO MEFs. MEFs, grown in DMEM plus 5%LPDS, were treated with 1 µM thapsigargin for 6 hr and subjected to real time PCR analysis. (c) TNFα expression in KO MEFs grown in <t>10%FBS</t> and treated with 1 µM thapsigargin for 6 hr and subjected to real time PCR analysis. Expression of each gene was normalized to GAPDH and reported as 2 −ΔΔCt . Relative mRNA level of WT untreated cells was assumed as 1. Results are given as mean ± s.d., (n = 7). *p
    Buffer Fbs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 88 stars, based on 42 article reviews
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    88
    Omega Scientific Inc cdt fbs
    GATA4 binding to DNA precedes ERα binding. U2OS-ERα cells were deprived of estrogen for 3 d in phenol red-free media containing 5% <t>CDT-FBS.</t> Cells were then treated for 0, 5, 15, 30, or 45 min with 10 n m E2. ChIP was performed with antibodies
    Cdt Fbs, supplied by Omega Scientific Inc, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Seradigm complete fbs
    GATA4 binding to DNA precedes ERα binding. U2OS-ERα cells were deprived of estrogen for 3 d in phenol red-free media containing 5% <t>CDT-FBS.</t> Cells were then treated for 0, 5, 15, 30, or 45 min with 10 n m E2. ChIP was performed with antibodies
    Complete Fbs, supplied by Seradigm, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.

    Journal: Biomedicines

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways

    doi: 10.3390/biomedicines6030073

    Figure Lengend Snippet: Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MCF-7 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.

    Article Snippet: Reagents Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA).

    Techniques: Expressing, Activation Assay, Incubation, Lysis, Protein Extraction, Western Blot, SDS Page

    Cell viability and cytotoxicity assessments of 4T1 cells treated with various CA-siRNA complexes involving ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs (1 nM) individually or in combination, for a period of 48 h. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for the next 48 h with the prepared complexes. MTT assay was performed and absorbance reading was taken at 595 nm with 630 nm as references wavelength. Data was presented as mean ± SD of triplicates.

    Journal: Biomedicines

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways

    doi: 10.3390/biomedicines6030073

    Figure Lengend Snippet: Cell viability and cytotoxicity assessments of 4T1 cells treated with various CA-siRNA complexes involving ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs (1 nM) individually or in combination, for a period of 48 h. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for the next 48 h with the prepared complexes. MTT assay was performed and absorbance reading was taken at 595 nm with 630 nm as references wavelength. Data was presented as mean ± SD of triplicates.

    Article Snippet: Reagents Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA).

    Techniques: Incubation, MTT Assay

    Cell viability and cytotoxicity assessments of MCF-7 cells ( A ), MDA-MB-231 ( B ) and 4T1 ( C ) cells treated with CA-ESR1 siRNA complexes for 48 h. Preparation of CA-ESR1 siRNA complexes involved introduction of different concentrations of ESR1 siRNA (0 to 10 nM) and 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4) medium. The mixture was allowed incubation at 37 °C for 30 min prior to addition of 10% FBS. The cells were incubated for the next 48 h with the prepared complexes. MTT assay was performed and absorbance reading was taken at 595 nm with 630 nm as references wavelength. Data was presented as mean ± SD of triplicates. * p

    Journal: Biomedicines

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways

    doi: 10.3390/biomedicines6030073

    Figure Lengend Snippet: Cell viability and cytotoxicity assessments of MCF-7 cells ( A ), MDA-MB-231 ( B ) and 4T1 ( C ) cells treated with CA-ESR1 siRNA complexes for 48 h. Preparation of CA-ESR1 siRNA complexes involved introduction of different concentrations of ESR1 siRNA (0 to 10 nM) and 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4) medium. The mixture was allowed incubation at 37 °C for 30 min prior to addition of 10% FBS. The cells were incubated for the next 48 h with the prepared complexes. MTT assay was performed and absorbance reading was taken at 595 nm with 630 nm as references wavelength. Data was presented as mean ± SD of triplicates. * p

    Article Snippet: Reagents Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA).

    Techniques: Multiple Displacement Amplification, Incubation, MTT Assay

    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in 4T1 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.

    Journal: Biomedicines

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways

    doi: 10.3390/biomedicines6030073

    Figure Lengend Snippet: Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in 4T1 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.

    Article Snippet: Reagents Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA).

    Techniques: Expressing, Activation Assay, Incubation, Lysis, Protein Extraction, Western Blot, SDS Page

    Cell viability and cytotoxicity assessments of MDA-MB-231 cells treated with various CA-siRNA complexes involving ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs (1 nM) individually or in combination, for a period of 48 h. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for the next 48 h with the prepared complexes. MTT assay was performed and absorbance reading was taken at 595 nm with 630 nm as references wavelength. Data was presented as mean ± SD of triplicates.

    Journal: Biomedicines

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways

    doi: 10.3390/biomedicines6030073

    Figure Lengend Snippet: Cell viability and cytotoxicity assessments of MDA-MB-231 cells treated with various CA-siRNA complexes involving ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs (1 nM) individually or in combination, for a period of 48 h. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for the next 48 h with the prepared complexes. MTT assay was performed and absorbance reading was taken at 595 nm with 630 nm as references wavelength. Data was presented as mean ± SD of triplicates.

    Article Snippet: Reagents Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA).

    Techniques: Multiple Displacement Amplification, Incubation, MTT Assay

    Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MDA-MB-231 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.

    Journal: Biomedicines

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways

    doi: 10.3390/biomedicines6030073

    Figure Lengend Snippet: Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MDA-MB-231 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.

    Article Snippet: Reagents Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA).

    Techniques: Expressing, Activation Assay, Multiple Displacement Amplification, Incubation, Lysis, Protein Extraction, Western Blot, SDS Page

    Cell viability and cytotoxicity assessments of MCF-7 cells treated with various CA-siRNA complexes involving ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs (1 nM) individually or in combination, for a period of 48 h. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for the next 48 h with the prepared complexes. MTT assay was performed and absorbance reading was taken at 595 nm with 630 nm as references wavelength. Data was presented as mean ± SD of triplicates.

    Journal: Biomedicines

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways

    doi: 10.3390/biomedicines6030073

    Figure Lengend Snippet: Cell viability and cytotoxicity assessments of MCF-7 cells treated with various CA-siRNA complexes involving ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs (1 nM) individually or in combination, for a period of 48 h. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl 2 to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for the next 48 h with the prepared complexes. MTT assay was performed and absorbance reading was taken at 595 nm with 630 nm as references wavelength. Data was presented as mean ± SD of triplicates.

    Article Snippet: Reagents Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA).

    Techniques: Incubation, MTT Assay

    qRT-PCR validation of the GEP microarray data ( blue ) in Sca1+ cells purified from BM or spleen of another set of HR-MDS mice ( two different hues of orange ), and CD34+ cells purified from MDS patients ( green ) compared to FBV/N or CD34 BM cells, respectively. Data expressed as fold change of expression of several genes of three pathways shown (angpt1 (signal transduction); cox4i1 and ndufv2 (oxidative metabolism); sdc1 (DNA processing)). qRT-PCR fold changes shown here are represented as the average of three experiments

    Journal: Journal of Hematology & Oncology

    Article Title: GEP analysis validates high risk MDS and acute myeloid leukemia post MDS mice models and highlights novel dysregulated pathways

    doi: 10.1186/s13045-016-0235-8

    Figure Lengend Snippet: qRT-PCR validation of the GEP microarray data ( blue ) in Sca1+ cells purified from BM or spleen of another set of HR-MDS mice ( two different hues of orange ), and CD34+ cells purified from MDS patients ( green ) compared to FBV/N or CD34 BM cells, respectively. Data expressed as fold change of expression of several genes of three pathways shown (angpt1 (signal transduction); cox4i1 and ndufv2 (oxidative metabolism); sdc1 (DNA processing)). qRT-PCR fold changes shown here are represented as the average of three experiments

    Article Snippet: Affymetrix exon array hybridization One microgram of total RNA extracted from Sca1+ spleen cells from the AML post MDS mice, HR-MDS mice, single-transgenic mice necessary to produce these AML post MDS and HR-MDS mice, (MRP8 NRASD12, MRP8 hBCL-2, tet hBCL-2) (called founder mice), and FBV/N control mice (n = 3 each) was labeled with Affymetrix reagents and hybridized to Affymetrix-GeneChip Mouse Exon 1.0 ST arrays.

    Techniques: Quantitative RT-PCR, Microarray, Purification, Mouse Assay, Expressing, Transduction

    Overexpression of 70z-Cbl promotes the loss of cell–cell contacts. (A) Parental MDCK cells, unstimulated or stimulated with 5 U/ml HGF for 24 h, as well as stable cell lines expressing c-Cbl (clone 8) and 70z-Cbl (clone 20), were grown on glass coverslips in DMEM containing 10% FBS. Cells were treated for 10 min with CSK buffer, fixed in 3.7% formaldehyde, and then labeled with anti-E-cadherin antibody followed by Cy3-conjugated anti-mouse antiserum (a, c, e, and g). Matching phase-contrast images are shown (b, d, f, and h). Photographs were taken at a magnification of ×63. (B) Whole cell lysates were separated on an 8% SDS-PAGE gel and immunoblotted with anti-E-cadherin antibody.

    Journal: Molecular Biology of the Cell

    Article Title: Cbl-transforming Variants Trigger a Cascade of Molecular Alterations That Lead to Epithelial Mesenchymal Conversion

    doi:

    Figure Lengend Snippet: Overexpression of 70z-Cbl promotes the loss of cell–cell contacts. (A) Parental MDCK cells, unstimulated or stimulated with 5 U/ml HGF for 24 h, as well as stable cell lines expressing c-Cbl (clone 8) and 70z-Cbl (clone 20), were grown on glass coverslips in DMEM containing 10% FBS. Cells were treated for 10 min with CSK buffer, fixed in 3.7% formaldehyde, and then labeled with anti-E-cadherin antibody followed by Cy3-conjugated anti-mouse antiserum (a, c, e, and g). Matching phase-contrast images are shown (b, d, f, and h). Photographs were taken at a magnification of ×63. (B) Whole cell lysates were separated on an 8% SDS-PAGE gel and immunoblotted with anti-E-cadherin antibody.

    Article Snippet: COS-1 and MDCK cells were maintained in DMEM containing 10% FBS (Life Technologies, Grand Island, NY).

    Techniques: Over Expression, Stable Transfection, Expressing, Labeling, SDS Page

    Tm7sf2 gene presides over an anti inflammatory loop. (a) Nuclear translocation of Nrf2 and NF-κB in WT and KO MEFs. MEFs, grown in DMEM plus 5%LPDS, were treated with 1 µM thapsigargin. At the indicated times, cells were collected and nuclear extracts subjected to Western blotting analyses with the indicated antibodies. Lamin B was used as loading control; (b) TNFα and HO-1 gene expression in WT and KO MEFs. MEFs, grown in DMEM plus 5%LPDS, were treated with 1 µM thapsigargin for 6 hr and subjected to real time PCR analysis. (c) TNFα expression in KO MEFs grown in 10%FBS and treated with 1 µM thapsigargin for 6 hr and subjected to real time PCR analysis. Expression of each gene was normalized to GAPDH and reported as 2 −ΔΔCt . Relative mRNA level of WT untreated cells was assumed as 1. Results are given as mean ± s.d., (n = 7). *p

    Journal: PLoS ONE

    Article Title: A Novel Role for Tm7sf2 Gene in Regulating TNF? Expression

    doi: 10.1371/journal.pone.0068017

    Figure Lengend Snippet: Tm7sf2 gene presides over an anti inflammatory loop. (a) Nuclear translocation of Nrf2 and NF-κB in WT and KO MEFs. MEFs, grown in DMEM plus 5%LPDS, were treated with 1 µM thapsigargin. At the indicated times, cells were collected and nuclear extracts subjected to Western blotting analyses with the indicated antibodies. Lamin B was used as loading control; (b) TNFα and HO-1 gene expression in WT and KO MEFs. MEFs, grown in DMEM plus 5%LPDS, were treated with 1 µM thapsigargin for 6 hr and subjected to real time PCR analysis. (c) TNFα expression in KO MEFs grown in 10%FBS and treated with 1 µM thapsigargin for 6 hr and subjected to real time PCR analysis. Expression of each gene was normalized to GAPDH and reported as 2 −ΔΔCt . Relative mRNA level of WT untreated cells was assumed as 1. Results are given as mean ± s.d., (n = 7). *p

    Article Snippet: MEFs were maintained in DMEM containing 10% FBS (Lonza, Milan, Italy) and switched to LPDS 24 hr prior to treatments.

    Techniques: Translocation Assay, Western Blot, Expressing, Real-time Polymerase Chain Reaction

    GATA4 binding to DNA precedes ERα binding. U2OS-ERα cells were deprived of estrogen for 3 d in phenol red-free media containing 5% CDT-FBS. Cells were then treated for 0, 5, 15, 30, or 45 min with 10 n m E2. ChIP was performed with antibodies

    Journal: Molecular Endocrinology

    Article Title: GATA4 Regulates Estrogen Receptor-?-Mediated Osteoblast Transcription

    doi: 10.1210/me.2010-0463

    Figure Lengend Snippet: GATA4 binding to DNA precedes ERα binding. U2OS-ERα cells were deprived of estrogen for 3 d in phenol red-free media containing 5% CDT-FBS. Cells were then treated for 0, 5, 15, 30, or 45 min with 10 n m E2. ChIP was performed with antibodies

    Article Snippet: Cells were hormone deprived by culture for 3 d in phenol red-free medium (Invitrogen) supplemented with 5% CDT-FBS (Omega Scientific, Tarzana, CA).

    Techniques: Binding Assay, Chromatin Immunoprecipitation

    GATA4 is expressed in osteoblasts. A, U2OS-ERα and MCF-7 cells were deprived of estrogen for 3 d in phenol red-free media containing 5% CDT-FBS. They were then treated with 10 n m E2 for 0, 3, 6, or 12 h, and RNA was obtained. GATA3 mRNA was analyzed

    Journal: Molecular Endocrinology

    Article Title: GATA4 Regulates Estrogen Receptor-?-Mediated Osteoblast Transcription

    doi: 10.1210/me.2010-0463

    Figure Lengend Snippet: GATA4 is expressed in osteoblasts. A, U2OS-ERα and MCF-7 cells were deprived of estrogen for 3 d in phenol red-free media containing 5% CDT-FBS. They were then treated with 10 n m E2 for 0, 3, 6, or 12 h, and RNA was obtained. GATA3 mRNA was analyzed

    Article Snippet: Cells were hormone deprived by culture for 3 d in phenol red-free medium (Invitrogen) supplemented with 5% CDT-FBS (Omega Scientific, Tarzana, CA).

    Techniques: