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PerkinElmer polymerase chain reaction
Polymerase Chain Reaction, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 80/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 80 stars, based on 60 article reviews
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polymerase chain reaction - by Bioz Stars, 2020-07
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Article Title: Cloning and expression of recombinant human pineal tryptophan hydroxylase in Escherichia coli: purification and characterization of the cloned enzyme.
Article Snippet: Reagents for the polymerase chain reaction were obtained from PerkinElmer (Norwalk, CT).

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    PerkinElmer neutralization assay hepg2 c3a cells
    Hepatit is E virus (HEV) infection kinetics in <t>HepG2/C3A</t> cells. ( A ) Immunofluorescence images show the expression of pORF2 (green) at different times after inoculation. The scale bar represents 50 μm. ( B ) The number of positive cells determined by immunofluorescence focus assay (IFA) is shown as a black line. The black dashed line represents average positive cells in the negative controls. HEV antigen pORF2 in the supernatant and cell lysate was detected by an HEV antigen test (red lines). The red dashed line (OD value = 0.17) represents the cut-off value of the negative controls. HEV RNA in the supernatants and cell lysates of HEV-infected HepG2 culture (blue lines). The blue short dashed line indicates the qRT-PCR detection limit. The results represent the means ± SEMs of four independent experiments.
    Neutralization Assay Hepg2 C3a Cells, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    PerkinElmer mitosox fluorescence
    The effect of GW9662 on ROS levels and IL-8 expression in H. pylori —infected AGS cells treated vs not treated with astaxanthin. The cells were treated with astaxanthin (5 μM) for 3 h without, or in combination with, the PPAR-γ antagonist GW9662 (5 μM), and then stimulated with H. pylori for 1 h (for intracellular and mitochondrial ROS levels), 4 h (for IL-8 mRNA level), and 24 h (for IL-8 protein level in the medium). ( A ) A plot of the relative intracellular ROS levels measured by DCF-DA fluorescence. ( B ) A plot of the relative mitochondrial ROS level measured by <t>MitoSOX</t> fluorescence. ( C ) A plot of the relative mRNA expression of IL-8 determined by real-time PCR analysis. ( D ) A plot of the concentration of IL-8 in the media determined by ELISA. * p
    Mitosox Fluorescence, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    PerkinElmer mcherry
    m 6 A depletion induces exon exclusion in synapse-associated genes. (A) Genes producing exon-excluded transcripts in P7 cKO cerebellums as compared with the Ctrl. (B) Enriched pathways of genes producing exon-excluded transcripts in P7 cKO cerebellums as compared with the Ctrl. Enriched pathways under the “Neuronal system” are expanded on the right. (C) Production of exon-excluded transcripts of major synapse-associated genes in P7 cKO cerebellums. Left, IGV tracks displaying the RNA-seq reads coverage in P7 Ctrl (blue) and cKO (red) cerebellum, the excluded exons are shaded with pink columns. Middle, calculated exon-exclusion percentage in P7 cKO cerebellums by the PSI value. Right, semiquantitative PCR detection of exon-excluded transcripts in P7 cKO cerebellums. (D) Images of the CGCs in the calcium detection assay. Grin1 001 or Grin1 011 was introduced into the cKO CGCs by electroporation and was co-expressed with <t>mCherry</t> (red). Fluo-4 (rainbow) indicated the cellular calcium concentration in the CGCs. Scale bar, 5 μm. (E) Statistical analysis of the cellular calcium concentration in the calcium detection assay. (F) CCK8 assay shows the survival rate of the Ctrl CGCs with overexpression of mCherry and cKO CGCs with overexpression of Grin1 001 or Grin1 011 in 3 DIV and 5 DIV. (G) Schematic diagram illustrating the mechanisms underlying the regulatory functions of METTL3-mediated m 6 A modification in cerebellar development. Red dots represent m 6 A modification. The box and line symbols in the right part of the diagram represent pre-spliced RNAs, with exons shown as boxes and introns shown as black lines. Exons in post-spliced RNAs are connected by red lines. Further information about this figure can be found in S1 Data . The data were represented as means ± SEM. For calcium concentration detection, n = 10; for CCK8 assay, n = 3. * p -value
    Mcherry, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer real time quantitative reverse transcription pcr rt pcr
    C. albicans cerebritis induces upregulation of amyloid precursor protein (APP). Mice were challenged with C. albicans as in Fig. 2 and RNA and protein extracted from whole brain. a <t>qRT-PCR</t> was performed to quantify <t>mRNA</t> for APP over 14 days. b Western blot analysis of APP over the same time period. c Densitometric analysis of the western blot data. d Immunofluorescence images of brain from C. albicans -infected wild-type mice 4 days post i.v. challenge with C. albicans showing staining for DAPI, IBA1, APP, NeuN, and merged images. Images are centered on fungal granulomas similar to those shown in Fig. 2 . e Repeat staining of fungal granulomas for DAPI, IBA1, APP, and calcofluor white (CW) comparing wild-type to app − / − mice. ( n = 4, mean ± S.E.M, * p
    Real Time Quantitative Reverse Transcription Pcr Rt Pcr, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Hepatit is E virus (HEV) infection kinetics in HepG2/C3A cells. ( A ) Immunofluorescence images show the expression of pORF2 (green) at different times after inoculation. The scale bar represents 50 μm. ( B ) The number of positive cells determined by immunofluorescence focus assay (IFA) is shown as a black line. The black dashed line represents average positive cells in the negative controls. HEV antigen pORF2 in the supernatant and cell lysate was detected by an HEV antigen test (red lines). The red dashed line (OD value = 0.17) represents the cut-off value of the negative controls. HEV RNA in the supernatants and cell lysates of HEV-infected HepG2 culture (blue lines). The blue short dashed line indicates the qRT-PCR detection limit. The results represent the means ± SEMs of four independent experiments.

    Journal: Viruses

    Article Title: An Optimized High-Throughput Neutralization Assay for Hepatitis E Virus (HEV) Involving Detection of Secreted Porf2

    doi: 10.3390/v11010064

    Figure Lengend Snippet: Hepatit is E virus (HEV) infection kinetics in HepG2/C3A cells. ( A ) Immunofluorescence images show the expression of pORF2 (green) at different times after inoculation. The scale bar represents 50 μm. ( B ) The number of positive cells determined by immunofluorescence focus assay (IFA) is shown as a black line. The black dashed line represents average positive cells in the negative controls. HEV antigen pORF2 in the supernatant and cell lysate was detected by an HEV antigen test (red lines). The red dashed line (OD value = 0.17) represents the cut-off value of the negative controls. HEV RNA in the supernatants and cell lysates of HEV-infected HepG2 culture (blue lines). The blue short dashed line indicates the qRT-PCR detection limit. The results represent the means ± SEMs of four independent experiments.

    Article Snippet: Neutralization Assay HepG2/C3A cells were seeded onto 96-well plates (PerkinElmer, Inc., Waltham, MA, USA) at a concentration of 3 × 104 cells per well a day before infection.

    Techniques: Infection, Immunofluorescence, Expressing, Quantitative RT-PCR

    The effect of GW9662 on ROS levels and IL-8 expression in H. pylori —infected AGS cells treated vs not treated with astaxanthin. The cells were treated with astaxanthin (5 μM) for 3 h without, or in combination with, the PPAR-γ antagonist GW9662 (5 μM), and then stimulated with H. pylori for 1 h (for intracellular and mitochondrial ROS levels), 4 h (for IL-8 mRNA level), and 24 h (for IL-8 protein level in the medium). ( A ) A plot of the relative intracellular ROS levels measured by DCF-DA fluorescence. ( B ) A plot of the relative mitochondrial ROS level measured by MitoSOX fluorescence. ( C ) A plot of the relative mRNA expression of IL-8 determined by real-time PCR analysis. ( D ) A plot of the concentration of IL-8 in the media determined by ELISA. * p

    Journal: Nutrients

    Article Title: Astaxanthin Inhibits Mitochondrial Dysfunction and Interleukin-8 Expression in Helicobacter pylori-Infected Gastric Epithelial Cells

    doi: 10.3390/nu10091320

    Figure Lengend Snippet: The effect of GW9662 on ROS levels and IL-8 expression in H. pylori —infected AGS cells treated vs not treated with astaxanthin. The cells were treated with astaxanthin (5 μM) for 3 h without, or in combination with, the PPAR-γ antagonist GW9662 (5 μM), and then stimulated with H. pylori for 1 h (for intracellular and mitochondrial ROS levels), 4 h (for IL-8 mRNA level), and 24 h (for IL-8 protein level in the medium). ( A ) A plot of the relative intracellular ROS levels measured by DCF-DA fluorescence. ( B ) A plot of the relative mitochondrial ROS level measured by MitoSOX fluorescence. ( C ) A plot of the relative mRNA expression of IL-8 determined by real-time PCR analysis. ( D ) A plot of the concentration of IL-8 in the media determined by ELISA. * p

    Article Snippet: The MitoSOX fluorescence was measured (excitation at 514 nm and emission at 585 nm) using a Victor5 multi-label counter (PerkinElmer Life and Analytical Sciences, Boston, MA, USA).

    Techniques: Expressing, Infection, Fluorescence, Real-time Polymerase Chain Reaction, Concentration Assay, Enzyme-linked Immunosorbent Assay

    Effect of apocynin on ROS levels and IL-8 expression and effect of astaxanthin on NADPH oxidase activity in H. pylori -infected AGS cells. The AGS cells were pre-treated with the indicated concentrations of a NADPH oxidase inhibitor apocynin for 3 h and then stimulated with H. pylori for 1 h (for intracellular and mitochondrial ROS levels), 4 h (for IL-8 mRNA level), and 24 h (for IL-8 protein level in the medium). ( A ) Plot of the relative ROS level in AGS cells measured by DCF-DA fluorescence. Column “None” corresponds to uninfected AGS cells, column “Control” to H. pylori -infected AGS cells, and columns “0.2” and “1” to H. pylori -infected AGS cells pretreated with 0.2 and 1 μM apocynin, respectively. ( B ) Plot of the relative mitochondrial ROS level in AGS cells measured by MitoSOX fluorescence. The description of the columns is the same as in ( A ). ( C ) mRNA expression of IL-8 was determined by real-time PCR analysis. The description of the columns is the same as in ( A ). ( D ) Plot of the relative concentration of IL-8 in the media of cultured AGS cells determined by using the ELISA method. The description of the columns is the same as in ( A ). ( E ) Plot of the relative NADPH oxidase activity in AGS cells. The cells were pre-treated with 5 μM astaxanthin for 3 h and then stimulated with H. pylori for 1 h. NADPH oxidase activity was measured by lucigenin assay. The description of the columns is the same as in ( A ). * p

    Journal: Nutrients

    Article Title: Astaxanthin Inhibits Mitochondrial Dysfunction and Interleukin-8 Expression in Helicobacter pylori-Infected Gastric Epithelial Cells

    doi: 10.3390/nu10091320

    Figure Lengend Snippet: Effect of apocynin on ROS levels and IL-8 expression and effect of astaxanthin on NADPH oxidase activity in H. pylori -infected AGS cells. The AGS cells were pre-treated with the indicated concentrations of a NADPH oxidase inhibitor apocynin for 3 h and then stimulated with H. pylori for 1 h (for intracellular and mitochondrial ROS levels), 4 h (for IL-8 mRNA level), and 24 h (for IL-8 protein level in the medium). ( A ) Plot of the relative ROS level in AGS cells measured by DCF-DA fluorescence. Column “None” corresponds to uninfected AGS cells, column “Control” to H. pylori -infected AGS cells, and columns “0.2” and “1” to H. pylori -infected AGS cells pretreated with 0.2 and 1 μM apocynin, respectively. ( B ) Plot of the relative mitochondrial ROS level in AGS cells measured by MitoSOX fluorescence. The description of the columns is the same as in ( A ). ( C ) mRNA expression of IL-8 was determined by real-time PCR analysis. The description of the columns is the same as in ( A ). ( D ) Plot of the relative concentration of IL-8 in the media of cultured AGS cells determined by using the ELISA method. The description of the columns is the same as in ( A ). ( E ) Plot of the relative NADPH oxidase activity in AGS cells. The cells were pre-treated with 5 μM astaxanthin for 3 h and then stimulated with H. pylori for 1 h. NADPH oxidase activity was measured by lucigenin assay. The description of the columns is the same as in ( A ). * p

    Article Snippet: The MitoSOX fluorescence was measured (excitation at 514 nm and emission at 585 nm) using a Victor5 multi-label counter (PerkinElmer Life and Analytical Sciences, Boston, MA, USA).

    Techniques: Expressing, Activity Assay, Infection, Fluorescence, Real-time Polymerase Chain Reaction, Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

    m 6 A depletion induces exon exclusion in synapse-associated genes. (A) Genes producing exon-excluded transcripts in P7 cKO cerebellums as compared with the Ctrl. (B) Enriched pathways of genes producing exon-excluded transcripts in P7 cKO cerebellums as compared with the Ctrl. Enriched pathways under the “Neuronal system” are expanded on the right. (C) Production of exon-excluded transcripts of major synapse-associated genes in P7 cKO cerebellums. Left, IGV tracks displaying the RNA-seq reads coverage in P7 Ctrl (blue) and cKO (red) cerebellum, the excluded exons are shaded with pink columns. Middle, calculated exon-exclusion percentage in P7 cKO cerebellums by the PSI value. Right, semiquantitative PCR detection of exon-excluded transcripts in P7 cKO cerebellums. (D) Images of the CGCs in the calcium detection assay. Grin1 001 or Grin1 011 was introduced into the cKO CGCs by electroporation and was co-expressed with mCherry (red). Fluo-4 (rainbow) indicated the cellular calcium concentration in the CGCs. Scale bar, 5 μm. (E) Statistical analysis of the cellular calcium concentration in the calcium detection assay. (F) CCK8 assay shows the survival rate of the Ctrl CGCs with overexpression of mCherry and cKO CGCs with overexpression of Grin1 001 or Grin1 011 in 3 DIV and 5 DIV. (G) Schematic diagram illustrating the mechanisms underlying the regulatory functions of METTL3-mediated m 6 A modification in cerebellar development. Red dots represent m 6 A modification. The box and line symbols in the right part of the diagram represent pre-spliced RNAs, with exons shown as boxes and introns shown as black lines. Exons in post-spliced RNAs are connected by red lines. Further information about this figure can be found in S1 Data . The data were represented as means ± SEM. For calcium concentration detection, n = 10; for CCK8 assay, n = 3. * p -value

    Journal: PLoS Biology

    Article Title: METTL3-mediated m6A modification is required for cerebellar development

    doi: 10.1371/journal.pbio.2004880

    Figure Lengend Snippet: m 6 A depletion induces exon exclusion in synapse-associated genes. (A) Genes producing exon-excluded transcripts in P7 cKO cerebellums as compared with the Ctrl. (B) Enriched pathways of genes producing exon-excluded transcripts in P7 cKO cerebellums as compared with the Ctrl. Enriched pathways under the “Neuronal system” are expanded on the right. (C) Production of exon-excluded transcripts of major synapse-associated genes in P7 cKO cerebellums. Left, IGV tracks displaying the RNA-seq reads coverage in P7 Ctrl (blue) and cKO (red) cerebellum, the excluded exons are shaded with pink columns. Middle, calculated exon-exclusion percentage in P7 cKO cerebellums by the PSI value. Right, semiquantitative PCR detection of exon-excluded transcripts in P7 cKO cerebellums. (D) Images of the CGCs in the calcium detection assay. Grin1 001 or Grin1 011 was introduced into the cKO CGCs by electroporation and was co-expressed with mCherry (red). Fluo-4 (rainbow) indicated the cellular calcium concentration in the CGCs. Scale bar, 5 μm. (E) Statistical analysis of the cellular calcium concentration in the calcium detection assay. (F) CCK8 assay shows the survival rate of the Ctrl CGCs with overexpression of mCherry and cKO CGCs with overexpression of Grin1 001 or Grin1 011 in 3 DIV and 5 DIV. (G) Schematic diagram illustrating the mechanisms underlying the regulatory functions of METTL3-mediated m 6 A modification in cerebellar development. Red dots represent m 6 A modification. The box and line symbols in the right part of the diagram represent pre-spliced RNAs, with exons shown as boxes and introns shown as black lines. Exons in post-spliced RNAs are connected by red lines. Further information about this figure can be found in S1 Data . The data were represented as means ± SEM. For calcium concentration detection, n = 10; for CCK8 assay, n = 3. * p -value

    Article Snippet: Fluo-4 (green) and mCherry (indicating the expression of appropriate plasmid, red) fluorescence were imaged with an inverted Perkin-Elmer microscope.

    Techniques: RNA Sequencing Assay, Polymerase Chain Reaction, Detection Assay, Electroporation, Concentration Assay, CCK-8 Assay, Over Expression, Modification

    C. albicans cerebritis induces upregulation of amyloid precursor protein (APP). Mice were challenged with C. albicans as in Fig. 2 and RNA and protein extracted from whole brain. a qRT-PCR was performed to quantify mRNA for APP over 14 days. b Western blot analysis of APP over the same time period. c Densitometric analysis of the western blot data. d Immunofluorescence images of brain from C. albicans -infected wild-type mice 4 days post i.v. challenge with C. albicans showing staining for DAPI, IBA1, APP, NeuN, and merged images. Images are centered on fungal granulomas similar to those shown in Fig. 2 . e Repeat staining of fungal granulomas for DAPI, IBA1, APP, and calcofluor white (CW) comparing wild-type to app − / − mice. ( n = 4, mean ± S.E.M, * p

    Journal: Nature Communications

    Article Title: Microglia and amyloid precursor protein coordinate control of transient Candida cerebritis with memory deficits

    doi: 10.1038/s41467-018-07991-4

    Figure Lengend Snippet: C. albicans cerebritis induces upregulation of amyloid precursor protein (APP). Mice were challenged with C. albicans as in Fig. 2 and RNA and protein extracted from whole brain. a qRT-PCR was performed to quantify mRNA for APP over 14 days. b Western blot analysis of APP over the same time period. c Densitometric analysis of the western blot data. d Immunofluorescence images of brain from C. albicans -infected wild-type mice 4 days post i.v. challenge with C. albicans showing staining for DAPI, IBA1, APP, NeuN, and merged images. Images are centered on fungal granulomas similar to those shown in Fig. 2 . e Repeat staining of fungal granulomas for DAPI, IBA1, APP, and calcofluor white (CW) comparing wild-type to app − / − mice. ( n = 4, mean ± S.E.M, * p

    Article Snippet: Relative expression of mRNA for APP was detected by two-step, real-time quantitative reverse transcription-PCR (RT-PCR) with the ABI Perkin Elmer Prism 5700 Sequence Detection System (Applied Biosystems, Foster City, CA) using Taqman probe (Mm00476361, Mm01344172, Invitrogen, Carlsbad, CA).

    Techniques: Mouse Assay, Quantitative RT-PCR, Western Blot, Immunofluorescence, Infection, Staining