droplet mda Search Results


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  • 99
    ATCC mda mb 231 cells
    Met-1 response to an AR agonist. (A) Met-1 cells were cultured in media containing 10% FBS. AR protein levels were measured by Western analysis. Mouse liver and testes were used as controls for mouse AR and the <t>MDA-MB-231,</t> SUM159PT and MDA-MB-453 cell lines were used to examine the relative levels of AR in Met-1 cells compared to human TNBC cells. α-Tubulin was used as a loading control. (B,C) Met-1 cells were grown in media with hormone-stripped serum (10% DCC) for 48 hours then treated with 0.1% EtOH or 10 nM Dihydrotestosterone (DHT) for 3 days. (B) Met-1 cells were formalin-fixed and paraffin embedded. 5 μm sections were stained for AR (1:800). Shown are representative images; 40× objective, scale bar = 50 μm; inset zoom ×2. The percentage of AR positive cells per field in five non-overlapping fields per sample; mean, ** p
    Mda Mb 231 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 4759 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher phi29 dna polymerase
    Size distribution of <t>DNA-magnesium-pyrophosphate</t> particles as a function of multiple displacement amplification reaction times, with and without the heat-inactivation step. At different time points (17, 38 and 66 h) of the MDA reaction in 3-pL droplets at 30 °C, the resulting DNA-Mg-PP i particles were released from droplets and imaged under TEM to measure their size. The first set of measurements (green) was made without heat-inactivation step. The second set of measurements (grey) was made with heat-inactivation step at 65 °C for 15 min that is typically used to inactivate <t>phi29</t> DNA polymerase and terminate the MDA reaction.
    Phi29 Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 365 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    ChemoCentryx cxcr7
    Depletion of <t>CXCR7</t> causes cell cycle arrest, but not apoptosis. wt-MCF7, MCF7-LTED, wt-SUM44 and SUM44-LTED cells were transfected with si control or si CXCR7 ± oestradiol (E2) for 72-hours. (A) Whole-cell extracts were immunoblotted and assessed for changes in expression of BCL2, cleaved poly(ADP-ribose) polymerase (PARP) and β-arrestins 1 and 2. (B) Changes in BCL2 were assessed by quantitative RT-PCR. Data are expressed relative to dextran-coated charcoal (DCC)–treated si control . (C) Assessment of alteration in S- and G 1 -phase accumulation as a result of CXCR7 depletion. Data are expressed as mean ± SEM. * P
    Cxcr7, supplied by ChemoCentryx, used in various techniques. Bioz Stars score: 92/100, based on 778 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Becton Dickinson type 1 collagen
    Depletion of <t>CXCR7</t> causes cell cycle arrest, but not apoptosis. wt-MCF7, MCF7-LTED, wt-SUM44 and SUM44-LTED cells were transfected with si control or si CXCR7 ± oestradiol (E2) for 72-hours. (A) Whole-cell extracts were immunoblotted and assessed for changes in expression of BCL2, cleaved poly(ADP-ribose) polymerase (PARP) and β-arrestins 1 and 2. (B) Changes in BCL2 were assessed by quantitative RT-PCR. Data are expressed relative to dextran-coated charcoal (DCC)–treated si control . (C) Assessment of alteration in S- and G 1 -phase accumulation as a result of CXCR7 depletion. Data are expressed as mean ± SEM. * P
    Type 1 Collagen, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 133 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen repli g single cell kit
    Depletion of <t>CXCR7</t> causes cell cycle arrest, but not apoptosis. wt-MCF7, MCF7-LTED, wt-SUM44 and SUM44-LTED cells were transfected with si control or si CXCR7 ± oestradiol (E2) for 72-hours. (A) Whole-cell extracts were immunoblotted and assessed for changes in expression of BCL2, cleaved poly(ADP-ribose) polymerase (PARP) and β-arrestins 1 and 2. (B) Changes in BCL2 were assessed by quantitative RT-PCR. Data are expressed relative to dextran-coated charcoal (DCC)–treated si control . (C) Assessment of alteration in S- and G 1 -phase accumulation as a result of CXCR7 depletion. Data are expressed as mean ± SEM. * P
    Repli G Single Cell Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 1527 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Illumina Inc 16s rrna fragments
    Single-cell sequencing of mouse gut microbes by ccSAG. SAGs from mouse gut microbes (n = 72) were obtained by single-droplet MDA, sequenced, and processed by ccSAG to obtain composite single-cell genomes. ( a ) Phylogenetic tree based on <t>16S</t> rDNA V3-V4. Phyla are highlighted in different colors. ( b ) Distribution of gut microbial phyla as determined from SAGs and metagenomic 16S rDNA. ( c ) Mean pairwise genomic similarity, as measured by BLAST. Strongly contaminated ( > 10%) samples or samples with no alignments were excluded from this analysis. ( d ) Phylogenetic tree of MGM1, MGM2, and mammal-associated Bacteroidales based on full length 16S <t>rRNA.</t> ( e ) Sequence mapping of cleaned reads of a putative polysaccharide lyase gene from single MGM1 cells (SAG04, SAG06, and SAG07), with the composite single-cell genome as reference. The composite genome is color-coded by base, and SNPs (A to C) in each read in SAG04, SAG06, and SAG07 are highlighted in corresponding base colors.
    16s Rrna Fragments, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 89/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher alexa488 conjugated phalloidin
    Single-cell sequencing of mouse gut microbes by ccSAG. SAGs from mouse gut microbes (n = 72) were obtained by single-droplet MDA, sequenced, and processed by ccSAG to obtain composite single-cell genomes. ( a ) Phylogenetic tree based on <t>16S</t> rDNA V3-V4. Phyla are highlighted in different colors. ( b ) Distribution of gut microbial phyla as determined from SAGs and metagenomic 16S rDNA. ( c ) Mean pairwise genomic similarity, as measured by BLAST. Strongly contaminated ( > 10%) samples or samples with no alignments were excluded from this analysis. ( d ) Phylogenetic tree of MGM1, MGM2, and mammal-associated Bacteroidales based on full length 16S <t>rRNA.</t> ( e ) Sequence mapping of cleaned reads of a putative polysaccharide lyase gene from single MGM1 cells (SAG04, SAG06, and SAG07), with the composite single-cell genome as reference. The composite genome is color-coded by base, and SNPs (A to C) in each read in SAG04, SAG06, and SAG07 are highlighted in corresponding base colors.
    Alexa488 Conjugated Phalloidin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 328 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ucp1  (Abcam)
    94
    Abcam ucp1
    Presence of cells with brown adipocyte phenotype and characteristics in HMLE HRASV12 xenografts over time. A–B, Xenografts at various stages of growth were fixed with glutaraldehyde, post-fixed with osmium tetroxide and embedded in Technovit. Low (A) and high (B) magnification pictures of hematoxylin/eosin-stained sections are shown. Cells (2×10 6 ) from a transplantable HMLE HRASV12 xenograft were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) in nude mice to develop xenografts, which were excised at various time points. C, Representative bright field images of xenografts at 1, 5, and 15 week. D, Western blot analysis of pooled lysates (n=3 mice/time point) from xenografts excised at various time points after growth subcutaneously in nude mice. A representative western blot analysis is shown (n=3). E, FACS analysis of PRDM16 (1- and 3 week xenograft), and <t>UCP1,</t> and UCP3 (1- and 5 week xenograft) labeled cells. Data shown is a representation of multiple experiments (n=3). F, Western blot analysis of key markers of angiogenesis (VEGF), proliferation (cyclin D1), and extracellular matrix proteins (ColA1, SMA and FN) in NT and T xenografts. G, Analysis of cellular bioenergetics of cells (4×10 4 ) obtained from 7-and 15-week xenografts (n=3,*p
    Ucp1, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 1844 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore phenol red free medium
    Presence of cells with brown adipocyte phenotype and characteristics in HMLE HRASV12 xenografts over time. A–B, Xenografts at various stages of growth were fixed with glutaraldehyde, post-fixed with osmium tetroxide and embedded in Technovit. Low (A) and high (B) magnification pictures of hematoxylin/eosin-stained sections are shown. Cells (2×10 6 ) from a transplantable HMLE HRASV12 xenograft were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) in nude mice to develop xenografts, which were excised at various time points. C, Representative bright field images of xenografts at 1, 5, and 15 week. D, Western blot analysis of pooled lysates (n=3 mice/time point) from xenografts excised at various time points after growth subcutaneously in nude mice. A representative western blot analysis is shown (n=3). E, FACS analysis of PRDM16 (1- and 3 week xenograft), and <t>UCP1,</t> and UCP3 (1- and 5 week xenograft) labeled cells. Data shown is a representation of multiple experiments (n=3). F, Western blot analysis of key markers of angiogenesis (VEGF), proliferation (cyclin D1), and extracellular matrix proteins (ColA1, SMA and FN) in NT and T xenografts. G, Analysis of cellular bioenergetics of cells (4×10 4 ) obtained from 7-and 15-week xenografts (n=3,*p
    Phenol Red Free Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 247 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher phenol red free imem
    Presence of cells with brown adipocyte phenotype and characteristics in HMLE HRASV12 xenografts over time. A–B, Xenografts at various stages of growth were fixed with glutaraldehyde, post-fixed with osmium tetroxide and embedded in Technovit. Low (A) and high (B) magnification pictures of hematoxylin/eosin-stained sections are shown. Cells (2×10 6 ) from a transplantable HMLE HRASV12 xenograft were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) in nude mice to develop xenografts, which were excised at various time points. C, Representative bright field images of xenografts at 1, 5, and 15 week. D, Western blot analysis of pooled lysates (n=3 mice/time point) from xenografts excised at various time points after growth subcutaneously in nude mice. A representative western blot analysis is shown (n=3). E, FACS analysis of PRDM16 (1- and 3 week xenograft), and <t>UCP1,</t> and UCP3 (1- and 5 week xenograft) labeled cells. Data shown is a representation of multiple experiments (n=3). F, Western blot analysis of key markers of angiogenesis (VEGF), proliferation (cyclin D1), and extracellular matrix proteins (ColA1, SMA and FN) in NT and T xenografts. G, Analysis of cellular bioenergetics of cells (4×10 4 ) obtained from 7-and 15-week xenografts (n=3,*p
    Phenol Red Free Imem, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Carl Zeiss zeiss lsm 780
    Presence of cells with brown adipocyte phenotype and characteristics in HMLE HRASV12 xenografts over time. A–B, Xenografts at various stages of growth were fixed with glutaraldehyde, post-fixed with osmium tetroxide and embedded in Technovit. Low (A) and high (B) magnification pictures of hematoxylin/eosin-stained sections are shown. Cells (2×10 6 ) from a transplantable HMLE HRASV12 xenograft were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) in nude mice to develop xenografts, which were excised at various time points. C, Representative bright field images of xenografts at 1, 5, and 15 week. D, Western blot analysis of pooled lysates (n=3 mice/time point) from xenografts excised at various time points after growth subcutaneously in nude mice. A representative western blot analysis is shown (n=3). E, FACS analysis of PRDM16 (1- and 3 week xenograft), and <t>UCP1,</t> and UCP3 (1- and 5 week xenograft) labeled cells. Data shown is a representation of multiple experiments (n=3). F, Western blot analysis of key markers of angiogenesis (VEGF), proliferation (cyclin D1), and extracellular matrix proteins (ColA1, SMA and FN) in NT and T xenografts. G, Analysis of cellular bioenergetics of cells (4×10 4 ) obtained from 7-and 15-week xenografts (n=3,*p
    Zeiss Lsm 780, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 94/100, based on 1749 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    TaKaRa lambda dna
    Presence of cells with brown adipocyte phenotype and characteristics in HMLE HRASV12 xenografts over time. A–B, Xenografts at various stages of growth were fixed with glutaraldehyde, post-fixed with osmium tetroxide and embedded in Technovit. Low (A) and high (B) magnification pictures of hematoxylin/eosin-stained sections are shown. Cells (2×10 6 ) from a transplantable HMLE HRASV12 xenograft were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) in nude mice to develop xenografts, which were excised at various time points. C, Representative bright field images of xenografts at 1, 5, and 15 week. D, Western blot analysis of pooled lysates (n=3 mice/time point) from xenografts excised at various time points after growth subcutaneously in nude mice. A representative western blot analysis is shown (n=3). E, FACS analysis of PRDM16 (1- and 3 week xenograft), and <t>UCP1,</t> and UCP3 (1- and 5 week xenograft) labeled cells. Data shown is a representation of multiple experiments (n=3). F, Western blot analysis of key markers of angiogenesis (VEGF), proliferation (cyclin D1), and extracellular matrix proteins (ColA1, SMA and FN) in NT and T xenografts. G, Analysis of cellular bioenergetics of cells (4×10 4 ) obtained from 7-and 15-week xenografts (n=3,*p
    Lambda Dna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 95/100, based on 119 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Illumina Inc gut microbiota
    Presence of cells with brown adipocyte phenotype and characteristics in HMLE HRASV12 xenografts over time. A–B, Xenografts at various stages of growth were fixed with glutaraldehyde, post-fixed with osmium tetroxide and embedded in Technovit. Low (A) and high (B) magnification pictures of hematoxylin/eosin-stained sections are shown. Cells (2×10 6 ) from a transplantable HMLE HRASV12 xenograft were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) in nude mice to develop xenografts, which were excised at various time points. C, Representative bright field images of xenografts at 1, 5, and 15 week. D, Western blot analysis of pooled lysates (n=3 mice/time point) from xenografts excised at various time points after growth subcutaneously in nude mice. A representative western blot analysis is shown (n=3). E, FACS analysis of PRDM16 (1- and 3 week xenograft), and <t>UCP1,</t> and UCP3 (1- and 5 week xenograft) labeled cells. Data shown is a representation of multiple experiments (n=3). F, Western blot analysis of key markers of angiogenesis (VEGF), proliferation (cyclin D1), and extracellular matrix proteins (ColA1, SMA and FN) in NT and T xenografts. G, Analysis of cellular bioenergetics of cells (4×10 4 ) obtained from 7-and 15-week xenografts (n=3,*p
    Gut Microbiota, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 93/100, based on 327 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore plasminogen free fibrinogen
    Presence of cells with brown adipocyte phenotype and characteristics in HMLE HRASV12 xenografts over time. A–B, Xenografts at various stages of growth were fixed with glutaraldehyde, post-fixed with osmium tetroxide and embedded in Technovit. Low (A) and high (B) magnification pictures of hematoxylin/eosin-stained sections are shown. Cells (2×10 6 ) from a transplantable HMLE HRASV12 xenograft were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) in nude mice to develop xenografts, which were excised at various time points. C, Representative bright field images of xenografts at 1, 5, and 15 week. D, Western blot analysis of pooled lysates (n=3 mice/time point) from xenografts excised at various time points after growth subcutaneously in nude mice. A representative western blot analysis is shown (n=3). E, FACS analysis of PRDM16 (1- and 3 week xenograft), and <t>UCP1,</t> and UCP3 (1- and 5 week xenograft) labeled cells. Data shown is a representation of multiple experiments (n=3). F, Western blot analysis of key markers of angiogenesis (VEGF), proliferation (cyclin D1), and extracellular matrix proteins (ColA1, SMA and FN) in NT and T xenografts. G, Analysis of cellular bioenergetics of cells (4×10 4 ) obtained from 7-and 15-week xenografts (n=3,*p
    Plasminogen Free Fibrinogen, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher platinum multiplex pcr master mix
    Demonstration of digital droplet MDA and its utility for nonspecific DNA quantification ( A ) Fluorescence microscopy images of droplets subjected to digital droplet MDA (ddMDA-upper row) and digital droplet <t>PCR</t> (ddPCR-lower row) for three concentrations of input material. Fluorescence was obtained using Eva Green (ddMDA) and <t>Taqman</t> probe (ddPCR). The disparity between digital MDA and PCR quantification corresponds to the nonspecific nature of MDA compared to specific PCR amplification ( B ) Fraction of observed versus predicted droplets. Fraction of fluorescent droplets is predicted assuming Poisson encapsulation of whole genomes. While ddPCR encapsulates one positive droplet per genome, ddMDA encapsulates one positive droplet per DNA segment. This enables nonspecific quantitation of nucleic acids and allows for the calculation of contamination and fragmentation of the sample.
    Platinum Multiplex Pcr Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 284 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore fluorescein isothiocyanate fitc
    Demonstration of digital droplet MDA and its utility for nonspecific DNA quantification ( A ) Fluorescence microscopy images of droplets subjected to digital droplet MDA (ddMDA-upper row) and digital droplet <t>PCR</t> (ddPCR-lower row) for three concentrations of input material. Fluorescence was obtained using Eva Green (ddMDA) and <t>Taqman</t> probe (ddPCR). The disparity between digital MDA and PCR quantification corresponds to the nonspecific nature of MDA compared to specific PCR amplification ( B ) Fraction of observed versus predicted droplets. Fraction of fluorescent droplets is predicted assuming Poisson encapsulation of whole genomes. While ddPCR encapsulates one positive droplet per genome, ddMDA encapsulates one positive droplet per DNA segment. This enables nonspecific quantitation of nucleic acids and allows for the calculation of contamination and fragmentation of the sample.
    Fluorescein Isothiocyanate Fitc, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3319 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    ATCC e coli k12
    Demonstration of digital droplet MDA and its utility for nonspecific DNA quantification ( A ) Fluorescence microscopy images of droplets subjected to digital droplet MDA (ddMDA-upper row) and digital droplet <t>PCR</t> (ddPCR-lower row) for three concentrations of input material. Fluorescence was obtained using Eva Green (ddMDA) and <t>Taqman</t> probe (ddPCR). The disparity between digital MDA and PCR quantification corresponds to the nonspecific nature of MDA compared to specific PCR amplification ( B ) Fraction of observed versus predicted droplets. Fraction of fluorescent droplets is predicted assuming Poisson encapsulation of whole genomes. While ddPCR encapsulates one positive droplet per genome, ddMDA encapsulates one positive droplet per DNA segment. This enables nonspecific quantitation of nucleic acids and allows for the calculation of contamination and fragmentation of the sample.
    E Coli K12, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 218 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore lipid droplet
    Demonstration of digital droplet MDA and its utility for nonspecific DNA quantification ( A ) Fluorescence microscopy images of droplets subjected to digital droplet MDA (ddMDA-upper row) and digital droplet <t>PCR</t> (ddPCR-lower row) for three concentrations of input material. Fluorescence was obtained using Eva Green (ddMDA) and <t>Taqman</t> probe (ddPCR). The disparity between digital MDA and PCR quantification corresponds to the nonspecific nature of MDA compared to specific PCR amplification ( B ) Fraction of observed versus predicted droplets. Fraction of fluorescent droplets is predicted assuming Poisson encapsulation of whole genomes. While ddPCR encapsulates one positive droplet per genome, ddMDA encapsulates one positive droplet per DNA segment. This enables nonspecific quantitation of nucleic acids and allows for the calculation of contamination and fragmentation of the sample.
    Lipid Droplet, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bio-Rad droplet digital polymerase chain reaction ddpcr
    Demonstration of digital droplet MDA and its utility for nonspecific DNA quantification ( A ) Fluorescence microscopy images of droplets subjected to digital droplet MDA (ddMDA-upper row) and digital droplet <t>PCR</t> (ddPCR-lower row) for three concentrations of input material. Fluorescence was obtained using Eva Green (ddMDA) and <t>Taqman</t> probe (ddPCR). The disparity between digital MDA and PCR quantification corresponds to the nonspecific nature of MDA compared to specific PCR amplification ( B ) Fraction of observed versus predicted droplets. Fraction of fluorescent droplets is predicted assuming Poisson encapsulation of whole genomes. While ddPCR encapsulates one positive droplet per genome, ddMDA encapsulates one positive droplet per DNA segment. This enables nonspecific quantitation of nucleic acids and allows for the calculation of contamination and fragmentation of the sample.
    Droplet Digital Polymerase Chain Reaction Ddpcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Biotium evagreen
    Droplet fusion and subsequent single-cell WGA in sd-MDA. ( a ) Histograms of droplets before (Cell lysate droplet: blue) and after fusion (SAG droplet: red). ( b ) Fluorescence image of droplets after the 1 st -round MDA reaction. E. coli cells were introduced at 0.1 cells/droplet and their genomes were amplified for 2 h with <t>Evagreen</t> dye. Scale bar; 100 μm. (c) Time-dependent appearance of the fluorescence signal during amplification of single E. coli genome. All data are presented as averaged intensities of fluorescent positive droplets measured with SD, and 100 droplets were analyzed at each time point. ( d ) Relationship between introduced E. coli cell concentration and the number of fluorescent positive droplets.
    Evagreen, supplied by Biotium, used in various techniques. Bioz Stars score: 94/100, based on 2149 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Illumina Inc illumina miseq system
    Droplet fusion and subsequent single-cell WGA in sd-MDA. ( a ) Histograms of droplets before (Cell lysate droplet: blue) and after fusion (SAG droplet: red). ( b ) Fluorescence image of droplets after the 1 st -round MDA reaction. E. coli cells were introduced at 0.1 cells/droplet and their genomes were amplified for 2 h with <t>Evagreen</t> dye. Scale bar; 100 μm. (c) Time-dependent appearance of the fluorescence signal during amplification of single E. coli genome. All data are presented as averaged intensities of fluorescent positive droplets measured with SD, and 100 droplets were analyzed at each time point. ( d ) Relationship between introduced E. coli cell concentration and the number of fluorescent positive droplets.
    Illumina Miseq System, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 99/100, based on 8008 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Droplet fusion and subsequent single-cell WGA in sd-MDA. ( a ) Histograms of droplets before (Cell lysate droplet: blue) and after fusion (SAG droplet: red). ( b ) Fluorescence image of droplets after the 1 st -round MDA reaction. E. coli cells were introduced at 0.1 cells/droplet and their genomes were amplified for 2 h with <t>Evagreen</t> dye. Scale bar; 100 μm. (c) Time-dependent appearance of the fluorescence signal during amplification of single E. coli genome. All data are presented as averaged intensities of fluorescent positive droplets measured with SD, and 100 droplets were analyzed at each time point. ( d ) Relationship between introduced E. coli cell concentration and the number of fluorescent positive droplets.
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    Droplet fusion and subsequent single-cell WGA in sd-MDA. ( a ) Histograms of droplets before (Cell lysate droplet: blue) and after fusion (SAG droplet: red). ( b ) Fluorescence image of droplets after the 1 st -round MDA reaction. E. coli cells were introduced at 0.1 cells/droplet and their genomes were amplified for 2 h with <t>Evagreen</t> dye. Scale bar; 100 μm. (c) Time-dependent appearance of the fluorescence signal during amplification of single E. coli genome. All data are presented as averaged intensities of fluorescent positive droplets measured with SD, and 100 droplets were analyzed at each time point. ( d ) Relationship between introduced E. coli cell concentration and the number of fluorescent positive droplets.
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    Image Search Results


    Met-1 response to an AR agonist. (A) Met-1 cells were cultured in media containing 10% FBS. AR protein levels were measured by Western analysis. Mouse liver and testes were used as controls for mouse AR and the MDA-MB-231, SUM159PT and MDA-MB-453 cell lines were used to examine the relative levels of AR in Met-1 cells compared to human TNBC cells. α-Tubulin was used as a loading control. (B,C) Met-1 cells were grown in media with hormone-stripped serum (10% DCC) for 48 hours then treated with 0.1% EtOH or 10 nM Dihydrotestosterone (DHT) for 3 days. (B) Met-1 cells were formalin-fixed and paraffin embedded. 5 μm sections were stained for AR (1:800). Shown are representative images; 40× objective, scale bar = 50 μm; inset zoom ×2. The percentage of AR positive cells per field in five non-overlapping fields per sample; mean, ** p

    Journal: Hormones & cancer

    Article Title: MMTV-PyMT and derived Met-1 mouse mammary tumor cells as models for studying the role of the androgen receptor in triple-negative breast cancer progression

    doi: 10.1007/s12672-017-0285-6

    Figure Lengend Snippet: Met-1 response to an AR agonist. (A) Met-1 cells were cultured in media containing 10% FBS. AR protein levels were measured by Western analysis. Mouse liver and testes were used as controls for mouse AR and the MDA-MB-231, SUM159PT and MDA-MB-453 cell lines were used to examine the relative levels of AR in Met-1 cells compared to human TNBC cells. α-Tubulin was used as a loading control. (B,C) Met-1 cells were grown in media with hormone-stripped serum (10% DCC) for 48 hours then treated with 0.1% EtOH or 10 nM Dihydrotestosterone (DHT) for 3 days. (B) Met-1 cells were formalin-fixed and paraffin embedded. 5 μm sections were stained for AR (1:800). Shown are representative images; 40× objective, scale bar = 50 μm; inset zoom ×2. The percentage of AR positive cells per field in five non-overlapping fields per sample; mean, ** p

    Article Snippet: MDA-MB-231 cells were purchased in 2008 from the American Type Culture Collection (ATCC, Rockville, MD) and maintained in MEM with 5% FBS, 1% non-essential amino acids and insulin.

    Techniques: Cell Culture, Western Blot, Multiple Displacement Amplification, Droplet Countercurrent Chromatography, Staining

    Size distribution of DNA-magnesium-pyrophosphate particles as a function of multiple displacement amplification reaction times, with and without the heat-inactivation step. At different time points (17, 38 and 66 h) of the MDA reaction in 3-pL droplets at 30 °C, the resulting DNA-Mg-PP i particles were released from droplets and imaged under TEM to measure their size. The first set of measurements (green) was made without heat-inactivation step. The second set of measurements (grey) was made with heat-inactivation step at 65 °C for 15 min that is typically used to inactivate phi29 DNA polymerase and terminate the MDA reaction.

    Journal: Micromachines

    Article Title: Droplet Microfluidics Approach for Single-DNA Molecule Amplification and Condensation into DNA-Magnesium-Pyrophosphate Particles

    doi: 10.3390/mi8020062

    Figure Lengend Snippet: Size distribution of DNA-magnesium-pyrophosphate particles as a function of multiple displacement amplification reaction times, with and without the heat-inactivation step. At different time points (17, 38 and 66 h) of the MDA reaction in 3-pL droplets at 30 °C, the resulting DNA-Mg-PP i particles were released from droplets and imaged under TEM to measure their size. The first set of measurements (green) was made without heat-inactivation step. The second set of measurements (grey) was made with heat-inactivation step at 65 °C for 15 min that is typically used to inactivate phi29 DNA polymerase and terminate the MDA reaction.

    Article Snippet: High droplet generation speed (~4000 s−1 ) allowed the collection of over 107 droplets in the form of an emulsion in less than 1 h. Droplets loaded with MDA reaction mix were collected off-chip into a collection tube and incubated for 16 h at 30 °C to initiate the isothermal DNA amplification reaction by phi29 DNA polymerase.

    Techniques: Multiple Displacement Amplification, Transmission Electron Microscopy

    Depletion of CXCR7 causes cell cycle arrest, but not apoptosis. wt-MCF7, MCF7-LTED, wt-SUM44 and SUM44-LTED cells were transfected with si control or si CXCR7 ± oestradiol (E2) for 72-hours. (A) Whole-cell extracts were immunoblotted and assessed for changes in expression of BCL2, cleaved poly(ADP-ribose) polymerase (PARP) and β-arrestins 1 and 2. (B) Changes in BCL2 were assessed by quantitative RT-PCR. Data are expressed relative to dextran-coated charcoal (DCC)–treated si control . (C) Assessment of alteration in S- and G 1 -phase accumulation as a result of CXCR7 depletion. Data are expressed as mean ± SEM. * P

    Journal: Breast Cancer Research : BCR

    Article Title: Identification of chemokine receptors as potential modulators of endocrine resistance in oestrogen receptor–positive breast cancers

    doi: 10.1186/s13058-014-0447-1

    Figure Lengend Snippet: Depletion of CXCR7 causes cell cycle arrest, but not apoptosis. wt-MCF7, MCF7-LTED, wt-SUM44 and SUM44-LTED cells were transfected with si control or si CXCR7 ± oestradiol (E2) for 72-hours. (A) Whole-cell extracts were immunoblotted and assessed for changes in expression of BCL2, cleaved poly(ADP-ribose) polymerase (PARP) and β-arrestins 1 and 2. (B) Changes in BCL2 were assessed by quantitative RT-PCR. Data are expressed relative to dextran-coated charcoal (DCC)–treated si control . (C) Assessment of alteration in S- and G 1 -phase accumulation as a result of CXCR7 depletion. Data are expressed as mean ± SEM. * P

    Article Snippet: Sections were stained for expression of CXCR7. (C) Wt-MCF7, MCF7-LTED and MDA MB 261 cells were stained for CXCR7 and expression visualised by FACS.

    Techniques: Transfection, Expressing, Quantitative RT-PCR, Droplet Countercurrent Chromatography

    CXCR4 , CXCR7, CXCL11 and CXCL12 expression in five oestrogen receptor–positive human breast cancer cell lines adapted to long-term oestrogen deprivation. Expression of CXCR4 (A) , CXCR7 (B) , CXCL11 (C) and CXCL12 (D) in wild-type (WT) and their corresponding long-term oestrogen-deprived (LTED) cell lines. CXCR, C-X-C chemokine receptor; CXCL, C-X-C chemokine ligand. Bars represent ± standard error of the mean (SEM) *P

    Journal: Breast Cancer Research : BCR

    Article Title: Identification of chemokine receptors as potential modulators of endocrine resistance in oestrogen receptor–positive breast cancers

    doi: 10.1186/s13058-014-0447-1

    Figure Lengend Snippet: CXCR4 , CXCR7, CXCL11 and CXCL12 expression in five oestrogen receptor–positive human breast cancer cell lines adapted to long-term oestrogen deprivation. Expression of CXCR4 (A) , CXCR7 (B) , CXCL11 (C) and CXCL12 (D) in wild-type (WT) and their corresponding long-term oestrogen-deprived (LTED) cell lines. CXCR, C-X-C chemokine receptor; CXCL, C-X-C chemokine ligand. Bars represent ± standard error of the mean (SEM) *P

    Article Snippet: Sections were stained for expression of CXCR7. (C) Wt-MCF7, MCF7-LTED and MDA MB 261 cells were stained for CXCR7 and expression visualised by FACS.

    Techniques: Expressing

    Analysis of CXCR4 and CXCR7 in cell lines and clinical data on association with recurrence of oestrogen receptor–positive breast cancer. wt-MCF7, MCF7-LTED, wt-SUM44 and SUM44-LTED cells were transfected with si CXCR4 (A) or si CXCR7 (B) versus si control ± exogenous oestradiol (E2) (1 nM). Cells were cultured for 6 days. Cell survival was measured using CellTiter-Glo. The data are expressed as fold changes relative to si control . Each treatment was carried out with eight replicates. The data shown are representative of a minimum of five independent experiments. Bars represent ± standard error of the mean (SEM). * P

    Journal: Breast Cancer Research : BCR

    Article Title: Identification of chemokine receptors as potential modulators of endocrine resistance in oestrogen receptor–positive breast cancers

    doi: 10.1186/s13058-014-0447-1

    Figure Lengend Snippet: Analysis of CXCR4 and CXCR7 in cell lines and clinical data on association with recurrence of oestrogen receptor–positive breast cancer. wt-MCF7, MCF7-LTED, wt-SUM44 and SUM44-LTED cells were transfected with si CXCR4 (A) or si CXCR7 (B) versus si control ± exogenous oestradiol (E2) (1 nM). Cells were cultured for 6 days. Cell survival was measured using CellTiter-Glo. The data are expressed as fold changes relative to si control . Each treatment was carried out with eight replicates. The data shown are representative of a minimum of five independent experiments. Bars represent ± standard error of the mean (SEM). * P

    Article Snippet: Sections were stained for expression of CXCR7. (C) Wt-MCF7, MCF7-LTED and MDA MB 261 cells were stained for CXCR7 and expression visualised by FACS.

    Techniques: Transfection, Cell Culture

    CXCR7 is required for the interaction between oestrogen receptor and PELP1. wt-MCF7 and MCF7-LTED cells were transfected with si control or si CXCR7 . (A) Oestrogen receptor/oestrogen response element (ER/ERE) transactivation was monitored with an ERE-linked luciferase reporter and pCH110 (β-galactosidase control) and expressed relative to dextran-coated charcoal (DCC) control. (B) Expression of TFF1 was assessed by quantitative RT-PCR, as previously described. Error bars represent ± SEM. * P

    Journal: Breast Cancer Research : BCR

    Article Title: Identification of chemokine receptors as potential modulators of endocrine resistance in oestrogen receptor–positive breast cancers

    doi: 10.1186/s13058-014-0447-1

    Figure Lengend Snippet: CXCR7 is required for the interaction between oestrogen receptor and PELP1. wt-MCF7 and MCF7-LTED cells were transfected with si control or si CXCR7 . (A) Oestrogen receptor/oestrogen response element (ER/ERE) transactivation was monitored with an ERE-linked luciferase reporter and pCH110 (β-galactosidase control) and expressed relative to dextran-coated charcoal (DCC) control. (B) Expression of TFF1 was assessed by quantitative RT-PCR, as previously described. Error bars represent ± SEM. * P

    Article Snippet: Sections were stained for expression of CXCR7. (C) Wt-MCF7, MCF7-LTED and MDA MB 261 cells were stained for CXCR7 and expression visualised by FACS.

    Techniques: Transfection, Luciferase, Droplet Countercurrent Chromatography, Expressing, Quantitative RT-PCR

    Schematic representation of signalling pathways of C-X-C chemokine receptors CXCR4 and CXCR7. MAPK, Mitogen-activated protein kinase; PKC, Protein kinase C.

    Journal: Breast Cancer Research : BCR

    Article Title: Identification of chemokine receptors as potential modulators of endocrine resistance in oestrogen receptor–positive breast cancers

    doi: 10.1186/s13058-014-0447-1

    Figure Lengend Snippet: Schematic representation of signalling pathways of C-X-C chemokine receptors CXCR4 and CXCR7. MAPK, Mitogen-activated protein kinase; PKC, Protein kinase C.

    Article Snippet: Sections were stained for expression of CXCR7. (C) Wt-MCF7, MCF7-LTED and MDA MB 261 cells were stained for CXCR7 and expression visualised by FACS.

    Techniques:

    Treatment of wt-MCF7 and MCF7-LTED cells with escalating concentrations of CXCR7 inhibitor CCX733 with or without exogenous oestradiol. wt-MCF7 cells (A) and MCF7-LTED cells (B) were treated with escalating concentrations of CCX733 for 6 days. Cell survival was measured using CellTiter-Glo. The data are expressed as fold changes relative to vehicle-treated control (0). (C) MCF7-LTED and wt-MCF7 cells were treated with CCX704 as a negative control. CXCR, Chemokine C-X-C receptor; DCC, Dextran-coated charcoal; E2, oestradiol; LTED, Long-term oestrogen deprivation; wt, Wild type. *P

    Journal: Breast Cancer Research : BCR

    Article Title: Identification of chemokine receptors as potential modulators of endocrine resistance in oestrogen receptor–positive breast cancers

    doi: 10.1186/s13058-014-0447-1

    Figure Lengend Snippet: Treatment of wt-MCF7 and MCF7-LTED cells with escalating concentrations of CXCR7 inhibitor CCX733 with or without exogenous oestradiol. wt-MCF7 cells (A) and MCF7-LTED cells (B) were treated with escalating concentrations of CCX733 for 6 days. Cell survival was measured using CellTiter-Glo. The data are expressed as fold changes relative to vehicle-treated control (0). (C) MCF7-LTED and wt-MCF7 cells were treated with CCX704 as a negative control. CXCR, Chemokine C-X-C receptor; DCC, Dextran-coated charcoal; E2, oestradiol; LTED, Long-term oestrogen deprivation; wt, Wild type. *P

    Article Snippet: Sections were stained for expression of CXCR7. (C) Wt-MCF7, MCF7-LTED and MDA MB 261 cells were stained for CXCR7 and expression visualised by FACS.

    Techniques: Negative Control, Droplet Countercurrent Chromatography

    Single-cell sequencing of mouse gut microbes by ccSAG. SAGs from mouse gut microbes (n = 72) were obtained by single-droplet MDA, sequenced, and processed by ccSAG to obtain composite single-cell genomes. ( a ) Phylogenetic tree based on 16S rDNA V3-V4. Phyla are highlighted in different colors. ( b ) Distribution of gut microbial phyla as determined from SAGs and metagenomic 16S rDNA. ( c ) Mean pairwise genomic similarity, as measured by BLAST. Strongly contaminated ( > 10%) samples or samples with no alignments were excluded from this analysis. ( d ) Phylogenetic tree of MGM1, MGM2, and mammal-associated Bacteroidales based on full length 16S rRNA. ( e ) Sequence mapping of cleaned reads of a putative polysaccharide lyase gene from single MGM1 cells (SAG04, SAG06, and SAG07), with the composite single-cell genome as reference. The composite genome is color-coded by base, and SNPs (A to C) in each read in SAG04, SAG06, and SAG07 are highlighted in corresponding base colors.

    Journal: Scientific Reports

    Article Title: Obtaining high-quality draft genomes from uncultured microbes by cleaning and co-assembly of single-cell amplified genomes

    doi: 10.1038/s41598-018-20384-3

    Figure Lengend Snippet: Single-cell sequencing of mouse gut microbes by ccSAG. SAGs from mouse gut microbes (n = 72) were obtained by single-droplet MDA, sequenced, and processed by ccSAG to obtain composite single-cell genomes. ( a ) Phylogenetic tree based on 16S rDNA V3-V4. Phyla are highlighted in different colors. ( b ) Distribution of gut microbial phyla as determined from SAGs and metagenomic 16S rDNA. ( c ) Mean pairwise genomic similarity, as measured by BLAST. Strongly contaminated ( > 10%) samples or samples with no alignments were excluded from this analysis. ( d ) Phylogenetic tree of MGM1, MGM2, and mammal-associated Bacteroidales based on full length 16S rRNA. ( e ) Sequence mapping of cleaned reads of a putative polysaccharide lyase gene from single MGM1 cells (SAG04, SAG06, and SAG07), with the composite single-cell genome as reference. The composite genome is color-coded by base, and SNPs (A to C) in each read in SAG04, SAG06, and SAG07 are highlighted in corresponding base colors.

    Article Snippet: To compare the phylogenetic distribution, 16S rRNA fragments (V3-V4) were also amplified from a metagenomic sample of gut microbiota and sequenced by MiSeq (Illumina, San Diego, CA, USA).

    Techniques: Sequencing, Multiple Displacement Amplification

    Presence of cells with brown adipocyte phenotype and characteristics in HMLE HRASV12 xenografts over time. A–B, Xenografts at various stages of growth were fixed with glutaraldehyde, post-fixed with osmium tetroxide and embedded in Technovit. Low (A) and high (B) magnification pictures of hematoxylin/eosin-stained sections are shown. Cells (2×10 6 ) from a transplantable HMLE HRASV12 xenograft were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) in nude mice to develop xenografts, which were excised at various time points. C, Representative bright field images of xenografts at 1, 5, and 15 week. D, Western blot analysis of pooled lysates (n=3 mice/time point) from xenografts excised at various time points after growth subcutaneously in nude mice. A representative western blot analysis is shown (n=3). E, FACS analysis of PRDM16 (1- and 3 week xenograft), and UCP1, and UCP3 (1- and 5 week xenograft) labeled cells. Data shown is a representation of multiple experiments (n=3). F, Western blot analysis of key markers of angiogenesis (VEGF), proliferation (cyclin D1), and extracellular matrix proteins (ColA1, SMA and FN) in NT and T xenografts. G, Analysis of cellular bioenergetics of cells (4×10 4 ) obtained from 7-and 15-week xenografts (n=3,*p

    Journal: Molecular cancer research : MCR

    Article Title: Increased expression of Beige/Brown adipose markers from host and breast cancer cells influence xenograft formation in mice

    doi: 10.1158/1541-7786.MCR-15-0151

    Figure Lengend Snippet: Presence of cells with brown adipocyte phenotype and characteristics in HMLE HRASV12 xenografts over time. A–B, Xenografts at various stages of growth were fixed with glutaraldehyde, post-fixed with osmium tetroxide and embedded in Technovit. Low (A) and high (B) magnification pictures of hematoxylin/eosin-stained sections are shown. Cells (2×10 6 ) from a transplantable HMLE HRASV12 xenograft were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) in nude mice to develop xenografts, which were excised at various time points. C, Representative bright field images of xenografts at 1, 5, and 15 week. D, Western blot analysis of pooled lysates (n=3 mice/time point) from xenografts excised at various time points after growth subcutaneously in nude mice. A representative western blot analysis is shown (n=3). E, FACS analysis of PRDM16 (1- and 3 week xenograft), and UCP1, and UCP3 (1- and 5 week xenograft) labeled cells. Data shown is a representation of multiple experiments (n=3). F, Western blot analysis of key markers of angiogenesis (VEGF), proliferation (cyclin D1), and extracellular matrix proteins (ColA1, SMA and FN) in NT and T xenografts. G, Analysis of cellular bioenergetics of cells (4×10 4 ) obtained from 7-and 15-week xenografts (n=3,*p

    Article Snippet: A, Sections of xenografts were analyzed for expression of UCP1 and extracellular matrix proteins (ColA1, Col 3, SMA and FN) by double immunofluorescence: B, Cells from dissociated xenograft (5 week) were analyzed for co-expression of UCP1 (Texas Red) and adiponectin (FITC) by double immunofluorescence and a representative picture is presented.

    Techniques: Staining, Mouse Assay, Western Blot, FACS, Labeling

    Depletion of UCP1 and Myf5 population reduced xenograft growth. A, Cells (2×10 6 ) from primary HMLE HRASV12 xenografts was mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorso-lateral) in nude mice (n=8). 5 weeks after implantation of cells, the mice were killed, tumors were excised and dissociated. Dissociated cells (2×10 7 ) from each of the eight xenografts were individually sorted by FACS into UCP1+ and UCP1− populations. From each of the xenografts, control unsorted (2×10 6 ) or UCP1 − (2×10 6 ) fraction of the sorted cells were injected subcutaneously (posterior dorsolateral) into nude mice (n=8). After 10 weeks, the mice were killed, tumors were excised and tumor weight was measured. B, Cells (2×10 6 ) from 7- and 15-week xenografts were injected into nude mice (subcutaneously, posterior dorsolateral) (n=3, per group per time point). Mice were killed, xenografts were excised and weighed at each of the time points. Data are presented as mean ± SD. (*p

    Journal: Molecular cancer research : MCR

    Article Title: Increased expression of Beige/Brown adipose markers from host and breast cancer cells influence xenograft formation in mice

    doi: 10.1158/1541-7786.MCR-15-0151

    Figure Lengend Snippet: Depletion of UCP1 and Myf5 population reduced xenograft growth. A, Cells (2×10 6 ) from primary HMLE HRASV12 xenografts was mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorso-lateral) in nude mice (n=8). 5 weeks after implantation of cells, the mice were killed, tumors were excised and dissociated. Dissociated cells (2×10 7 ) from each of the eight xenografts were individually sorted by FACS into UCP1+ and UCP1− populations. From each of the xenografts, control unsorted (2×10 6 ) or UCP1 − (2×10 6 ) fraction of the sorted cells were injected subcutaneously (posterior dorsolateral) into nude mice (n=8). After 10 weeks, the mice were killed, tumors were excised and tumor weight was measured. B, Cells (2×10 6 ) from 7- and 15-week xenografts were injected into nude mice (subcutaneously, posterior dorsolateral) (n=3, per group per time point). Mice were killed, xenografts were excised and weighed at each of the time points. Data are presented as mean ± SD. (*p

    Article Snippet: A, Sections of xenografts were analyzed for expression of UCP1 and extracellular matrix proteins (ColA1, Col 3, SMA and FN) by double immunofluorescence: B, Cells from dissociated xenograft (5 week) were analyzed for co-expression of UCP1 (Texas Red) and adiponectin (FITC) by double immunofluorescence and a representative picture is presented.

    Techniques: Mouse Assay, FACS, Injection

    . E. Paraffin embedded sections of human breast tumors was immunolabeled for UCP1 and representative images of estrogen receptor positive (ER + ) and negative (ER − ) tumor sections are shown. F, A representative model suggesting the role of UCP1 + /PRDM16 + population during tumor development.

    Journal: Molecular cancer research : MCR

    Article Title: Increased expression of Beige/Brown adipose markers from host and breast cancer cells influence xenograft formation in mice

    doi: 10.1158/1541-7786.MCR-15-0151

    Figure Lengend Snippet: . E. Paraffin embedded sections of human breast tumors was immunolabeled for UCP1 and representative images of estrogen receptor positive (ER + ) and negative (ER − ) tumor sections are shown. F, A representative model suggesting the role of UCP1 + /PRDM16 + population during tumor development.

    Article Snippet: A, Sections of xenografts were analyzed for expression of UCP1 and extracellular matrix proteins (ColA1, Col 3, SMA and FN) by double immunofluorescence: B, Cells from dissociated xenograft (5 week) were analyzed for co-expression of UCP1 (Texas Red) and adiponectin (FITC) by double immunofluorescence and a representative picture is presented.

    Techniques: Immunolabeling

    Transplantable HMLE HRASV12 xenografts show multi-locular lipid droplets and express beige/brown adipocyte-specific proteins. Cells (2×10 6 ) from primary HMLE HRASV12 xenografts were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) to develop xenografts. A–C, Low and high magnification images of hematoxylin/eosin-stained sections of transplantable (T) xenografts (7–10 week). Representative data is shown from multiple experiments (n=6). D–E, Bright field low and high magnification images of T xenografts (5–7 week) that were fixed in glutaraldehyde, post-fixed with osmium tetroxide, and stained with toluidine blue, showing brown stained lipid droplets. A representative image is shown from multiple experiments (n=6). F, Immuno-labelling of paraffin embedded sections of T xenograft for UCP1 protein. G–H, Relative gene expression of beige/brown adipocyte markers from T and NT xenografts (5 week) using human-specific primers (*p

    Journal: Molecular cancer research : MCR

    Article Title: Increased expression of Beige/Brown adipose markers from host and breast cancer cells influence xenograft formation in mice

    doi: 10.1158/1541-7786.MCR-15-0151

    Figure Lengend Snippet: Transplantable HMLE HRASV12 xenografts show multi-locular lipid droplets and express beige/brown adipocyte-specific proteins. Cells (2×10 6 ) from primary HMLE HRASV12 xenografts were mixed with matrigel (1:1) and implanted at a subcutaneous site (posterior dorsolateral) to develop xenografts. A–C, Low and high magnification images of hematoxylin/eosin-stained sections of transplantable (T) xenografts (7–10 week). Representative data is shown from multiple experiments (n=6). D–E, Bright field low and high magnification images of T xenografts (5–7 week) that were fixed in glutaraldehyde, post-fixed with osmium tetroxide, and stained with toluidine blue, showing brown stained lipid droplets. A representative image is shown from multiple experiments (n=6). F, Immuno-labelling of paraffin embedded sections of T xenograft for UCP1 protein. G–H, Relative gene expression of beige/brown adipocyte markers from T and NT xenografts (5 week) using human-specific primers (*p

    Article Snippet: A, Sections of xenografts were analyzed for expression of UCP1 and extracellular matrix proteins (ColA1, Col 3, SMA and FN) by double immunofluorescence: B, Cells from dissociated xenograft (5 week) were analyzed for co-expression of UCP1 (Texas Red) and adiponectin (FITC) by double immunofluorescence and a representative picture is presented.

    Techniques: Staining, Expressing

    Demonstration of digital droplet MDA and its utility for nonspecific DNA quantification ( A ) Fluorescence microscopy images of droplets subjected to digital droplet MDA (ddMDA-upper row) and digital droplet PCR (ddPCR-lower row) for three concentrations of input material. Fluorescence was obtained using Eva Green (ddMDA) and Taqman probe (ddPCR). The disparity between digital MDA and PCR quantification corresponds to the nonspecific nature of MDA compared to specific PCR amplification ( B ) Fraction of observed versus predicted droplets. Fraction of fluorescent droplets is predicted assuming Poisson encapsulation of whole genomes. While ddPCR encapsulates one positive droplet per genome, ddMDA encapsulates one positive droplet per DNA segment. This enables nonspecific quantitation of nucleic acids and allows for the calculation of contamination and fragmentation of the sample.

    Journal: Nucleic Acids Research

    Article Title: Enhanced sequencing coverage with digital droplet multiple displacement amplification

    doi: 10.1093/nar/gkv1493

    Figure Lengend Snippet: Demonstration of digital droplet MDA and its utility for nonspecific DNA quantification ( A ) Fluorescence microscopy images of droplets subjected to digital droplet MDA (ddMDA-upper row) and digital droplet PCR (ddPCR-lower row) for three concentrations of input material. Fluorescence was obtained using Eva Green (ddMDA) and Taqman probe (ddPCR). The disparity between digital MDA and PCR quantification corresponds to the nonspecific nature of MDA compared to specific PCR amplification ( B ) Fraction of observed versus predicted droplets. Fraction of fluorescent droplets is predicted assuming Poisson encapsulation of whole genomes. While ddPCR encapsulates one positive droplet per genome, ddMDA encapsulates one positive droplet per DNA segment. This enables nonspecific quantitation of nucleic acids and allows for the calculation of contamination and fragmentation of the sample.

    Article Snippet: For digital PCR, the template is mixed in bulk with primers (IDT), TaqMan probe (IDT) and 2X Platinum Multiplex PCR Master Mix (Life Technologies, catalog no. 4464268) in a total volume of 100 μl.

    Techniques: Multiple Displacement Amplification, Fluorescence, Microscopy, Polymerase Chain Reaction, Amplification, Quantitation Assay

    Droplet fusion and subsequent single-cell WGA in sd-MDA. ( a ) Histograms of droplets before (Cell lysate droplet: blue) and after fusion (SAG droplet: red). ( b ) Fluorescence image of droplets after the 1 st -round MDA reaction. E. coli cells were introduced at 0.1 cells/droplet and their genomes were amplified for 2 h with Evagreen dye. Scale bar; 100 μm. (c) Time-dependent appearance of the fluorescence signal during amplification of single E. coli genome. All data are presented as averaged intensities of fluorescent positive droplets measured with SD, and 100 droplets were analyzed at each time point. ( d ) Relationship between introduced E. coli cell concentration and the number of fluorescent positive droplets.

    Journal: Scientific Reports

    Article Title: Massively parallel whole genome amplification for single-cell sequencing using droplet microfluidics

    doi: 10.1038/s41598-017-05436-4

    Figure Lengend Snippet: Droplet fusion and subsequent single-cell WGA in sd-MDA. ( a ) Histograms of droplets before (Cell lysate droplet: blue) and after fusion (SAG droplet: red). ( b ) Fluorescence image of droplets after the 1 st -round MDA reaction. E. coli cells were introduced at 0.1 cells/droplet and their genomes were amplified for 2 h with Evagreen dye. Scale bar; 100 μm. (c) Time-dependent appearance of the fluorescence signal during amplification of single E. coli genome. All data are presented as averaged intensities of fluorescent positive droplets measured with SD, and 100 droplets were analyzed at each time point. ( d ) Relationship between introduced E. coli cell concentration and the number of fluorescent positive droplets.

    Article Snippet: Droplet fusion and single-cell whole genome amplification Prior to reagent introduction into the device, an MDA mixture (REPLI-g Single Cell Kit, QIAGEN) was prepared containing 1.5 μL of stop buffer, 1.5 μL of H2 O, 14.5 μL of reaction buffer, 1.0 μL of enzyme mix, 2.5 μL of 10% Tween-20 (1% v/v concentration), and 0.5 μL of 20x Evagreen (Biotium, Fremont, CA, USA), for use in a 21.5-μL reaction volume.

    Techniques: Whole Genome Amplification, Multiple Displacement Amplification, Fluorescence, Amplification, Concentration Assay