Structured Review

Sony sh800z cell sorter
C8 augments important maturation markers specifically in INS-producing β cells. Quantitative RT-PCR analysis was performed to measure the relative expression levels of the β cell lineage markers INS , UCN3 , and GCK and the transcription factors PDX1 , NKX6.1 , and MAFA from sorted Venus+ and NC cells with or without treatment with C8 and/or bFGF. Briefly, #9–15 hiPSCs were differentiated as described above and treated with 5 μM C8 or 5 μM C8 and 50 ng/mL bFGF in the third stage of differentiation. After differentiation, samples were dissociated with TrypLE and sorted with an <t>SH800Z</t> cell sorter (Sony) for Venus+ and NC cells. Fifty cells from each group were sorted into 5 μL FCP reagent, which is a lysate reagent of the QIAGEN Fast Lane cDNA kit. For cDNA synthesis, 2 μL of the samples were used, with 0.5 μL of the 10 μL cDNA samples used for quantitative RT-PCR. Error bars indicate SD, n = 3. *p
Sh800z Cell Sorter, supplied by Sony, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/sh800z cell sorter/product/Sony
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sh800z cell sorter - by Bioz Stars, 2021-07
86/100 stars

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1) Product Images from "An inhibitor of fibroblast growth factor receptor-1 (FGFR1) promotes late-stage terminal differentiation from NGN3+ pancreatic endocrine progenitors"

Article Title: An inhibitor of fibroblast growth factor receptor-1 (FGFR1) promotes late-stage terminal differentiation from NGN3+ pancreatic endocrine progenitors

Journal: Scientific Reports

doi: 10.1038/srep35908

C8 augments important maturation markers specifically in INS-producing β cells. Quantitative RT-PCR analysis was performed to measure the relative expression levels of the β cell lineage markers INS , UCN3 , and GCK and the transcription factors PDX1 , NKX6.1 , and MAFA from sorted Venus+ and NC cells with or without treatment with C8 and/or bFGF. Briefly, #9–15 hiPSCs were differentiated as described above and treated with 5 μM C8 or 5 μM C8 and 50 ng/mL bFGF in the third stage of differentiation. After differentiation, samples were dissociated with TrypLE and sorted with an SH800Z cell sorter (Sony) for Venus+ and NC cells. Fifty cells from each group were sorted into 5 μL FCP reagent, which is a lysate reagent of the QIAGEN Fast Lane cDNA kit. For cDNA synthesis, 2 μL of the samples were used, with 0.5 μL of the 10 μL cDNA samples used for quantitative RT-PCR. Error bars indicate SD, n = 3. *p
Figure Legend Snippet: C8 augments important maturation markers specifically in INS-producing β cells. Quantitative RT-PCR analysis was performed to measure the relative expression levels of the β cell lineage markers INS , UCN3 , and GCK and the transcription factors PDX1 , NKX6.1 , and MAFA from sorted Venus+ and NC cells with or without treatment with C8 and/or bFGF. Briefly, #9–15 hiPSCs were differentiated as described above and treated with 5 μM C8 or 5 μM C8 and 50 ng/mL bFGF in the third stage of differentiation. After differentiation, samples were dissociated with TrypLE and sorted with an SH800Z cell sorter (Sony) for Venus+ and NC cells. Fifty cells from each group were sorted into 5 μL FCP reagent, which is a lysate reagent of the QIAGEN Fast Lane cDNA kit. For cDNA synthesis, 2 μL of the samples were used, with 0.5 μL of the 10 μL cDNA samples used for quantitative RT-PCR. Error bars indicate SD, n = 3. *p

Techniques Used: Quantitative RT-PCR, Expressing

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Co-Culture Assay:

Article Title: Ninjurin1 Plays a Crucial Role in Pulmonary Fibrosis by Promoting Interaction between Macrophages and Alveolar Epithelial Cells
Article Snippet: Co-culture of MLE-12 and Raw264.7 WT or Ninj1 KO MLE-12 cells with or without BLM (50 µg/ml) treatment for 12 hours and CFSE-stained WT or Ninj1 KO Raw264.7 cells were co-cultured for 30 min, and the number of Raw264.7 cells bound to MLE-12 cells was assessed using the BD FACSCalibur (BD Bioscience). .. For RNA and protein analysis, after co-culture of WT or Ninj1 KO MLE-12 and CFSE-stained WT or Ninj1 KO Raw264.7 for 6 hours, Raw264.7 cells were sorted using the SONY cell sorter SH800Z (SONY, Tokyo, Japan), and RNA and protein were isolated from the sorted Raw264.7 cells. .. To collect conditioned media (CM), the media was changed to serum-free medium after 6 hours of co-culture and CM was collected 12 hours after media change.

Isolation:

Article Title: Ninjurin1 Plays a Crucial Role in Pulmonary Fibrosis by Promoting Interaction between Macrophages and Alveolar Epithelial Cells
Article Snippet: Co-culture of MLE-12 and Raw264.7 WT or Ninj1 KO MLE-12 cells with or without BLM (50 µg/ml) treatment for 12 hours and CFSE-stained WT or Ninj1 KO Raw264.7 cells were co-cultured for 30 min, and the number of Raw264.7 cells bound to MLE-12 cells was assessed using the BD FACSCalibur (BD Bioscience). .. For RNA and protein analysis, after co-culture of WT or Ninj1 KO MLE-12 and CFSE-stained WT or Ninj1 KO Raw264.7 for 6 hours, Raw264.7 cells were sorted using the SONY cell sorter SH800Z (SONY, Tokyo, Japan), and RNA and protein were isolated from the sorted Raw264.7 cells. .. To collect conditioned media (CM), the media was changed to serum-free medium after 6 hours of co-culture and CM was collected 12 hours after media change.

Article Title: Dynamic MAPK/ERK Activity Sustains Nephron Progenitors through Niche Regulation and Primes Precursors for Differentiation
Article Snippet: Quantification of NP perimeter, roundness, contact surface length, and angle toward UB epithelium was obtained from paraffin sections stained with CTNND1, CALB1, and Hoechst. .. Six2-TGC tg/+ -containing control (n = 34) and dko (n = 22) E13.5 kidneys were dissociated into single-cell suspensions with 0.25% trypsin, and GFP+ NP cells were isolated using fluorescence-activated cell sorting (FACS) with a Sony SH800Z cell sorter. .. Cells were subsequently fixed with 70% EtOH, treated with RNAse A (Thermo Fisher Scientific), and stained with propidium iodide (Invitrogen).

Expressing:

Article Title: A screening system for identifying interacting proteins using biomolecular fluorescence complementation and transposon gene trap
Article Snippet: Cell culture HeLa and HEK293T were maintained in DMEM (Fujifilm WAKO, Tokyo, Japan) supplemented with 10% FBS (Nichirei Biosciences, Tokyo, Japan), 100 μg/mL streptomycin sulfate, 100 U/mL penicillin G potassium (SMPG) at 37°C under 5% CO2. .. Bait cell line After infecting HeLa cells with a retroviral expression vector (pMXs-p65-mKGN-IRES-blaR), the cells were selected in bulk with blasticidin and then single cell sorting was performed using a cell sorter SH800Z (Sony, Tokyo, Japan). .. A cell clone that was able to withstand repeated single cell sorting, had high protein expression of p65-mKGN, and high BiFC fluorescence with the introduction of p50-mKGC was selected.

Article Title: Tracing the transitions from pluripotency to germ cell fate with CRISPR screening
Article Snippet: 100 ng of total RNA from triplicate biological independent experiments was used as input for the NEBNext Ultra RNA library Prep Kit for Illumina® (NEB), with the library generated as per manufacturer’s instructions. .. RNA-seq and gene expression For RNA-seq of Nr5a2 −/− , Zfp296 −/− and matched WT control cell lines, EpiLC, d2 or d6 PGCLC were dissociated into single-cell solutions with TrypLE and sorted with a Sony SH800Z or a MoFlow high-speed cell sorter (Beckman Coulter) based on appropriate Stella -GFP and Esg1 -tdTomato expression. .. Samples were collected in 150 μl of extraction buffer from the PicoPure RNA isolation kit (Life Technologies) and rapidly frozen on dry ice or liquid nitrogen.

Plasmid Preparation:

Article Title: A screening system for identifying interacting proteins using biomolecular fluorescence complementation and transposon gene trap
Article Snippet: Cell culture HeLa and HEK293T were maintained in DMEM (Fujifilm WAKO, Tokyo, Japan) supplemented with 10% FBS (Nichirei Biosciences, Tokyo, Japan), 100 μg/mL streptomycin sulfate, 100 U/mL penicillin G potassium (SMPG) at 37°C under 5% CO2. .. Bait cell line After infecting HeLa cells with a retroviral expression vector (pMXs-p65-mKGN-IRES-blaR), the cells were selected in bulk with blasticidin and then single cell sorting was performed using a cell sorter SH800Z (Sony, Tokyo, Japan). .. A cell clone that was able to withstand repeated single cell sorting, had high protein expression of p65-mKGN, and high BiFC fluorescence with the introduction of p50-mKGC was selected.

FACS:

Article Title: A screening system for identifying interacting proteins using biomolecular fluorescence complementation and transposon gene trap
Article Snippet: Cell culture HeLa and HEK293T were maintained in DMEM (Fujifilm WAKO, Tokyo, Japan) supplemented with 10% FBS (Nichirei Biosciences, Tokyo, Japan), 100 μg/mL streptomycin sulfate, 100 U/mL penicillin G potassium (SMPG) at 37°C under 5% CO2. .. Bait cell line After infecting HeLa cells with a retroviral expression vector (pMXs-p65-mKGN-IRES-blaR), the cells were selected in bulk with blasticidin and then single cell sorting was performed using a cell sorter SH800Z (Sony, Tokyo, Japan). .. A cell clone that was able to withstand repeated single cell sorting, had high protein expression of p65-mKGN, and high BiFC fluorescence with the introduction of p50-mKGC was selected.

Article Title: Dynamic MAPK/ERK Activity Sustains Nephron Progenitors through Niche Regulation and Primes Precursors for Differentiation
Article Snippet: Quantification of NP perimeter, roundness, contact surface length, and angle toward UB epithelium was obtained from paraffin sections stained with CTNND1, CALB1, and Hoechst. .. Six2-TGC tg/+ -containing control (n = 34) and dko (n = 22) E13.5 kidneys were dissociated into single-cell suspensions with 0.25% trypsin, and GFP+ NP cells were isolated using fluorescence-activated cell sorting (FACS) with a Sony SH800Z cell sorter. .. Cells were subsequently fixed with 70% EtOH, treated with RNAse A (Thermo Fisher Scientific), and stained with propidium iodide (Invitrogen).

Fluorescence:

Article Title: Dynamic MAPK/ERK Activity Sustains Nephron Progenitors through Niche Regulation and Primes Precursors for Differentiation
Article Snippet: Quantification of NP perimeter, roundness, contact surface length, and angle toward UB epithelium was obtained from paraffin sections stained with CTNND1, CALB1, and Hoechst. .. Six2-TGC tg/+ -containing control (n = 34) and dko (n = 22) E13.5 kidneys were dissociated into single-cell suspensions with 0.25% trypsin, and GFP+ NP cells were isolated using fluorescence-activated cell sorting (FACS) with a Sony SH800Z cell sorter. .. Cells were subsequently fixed with 70% EtOH, treated with RNAse A (Thermo Fisher Scientific), and stained with propidium iodide (Invitrogen).

RNA Sequencing Assay:

Article Title: Tracing the transitions from pluripotency to germ cell fate with CRISPR screening
Article Snippet: 100 ng of total RNA from triplicate biological independent experiments was used as input for the NEBNext Ultra RNA library Prep Kit for Illumina® (NEB), with the library generated as per manufacturer’s instructions. .. RNA-seq and gene expression For RNA-seq of Nr5a2 −/− , Zfp296 −/− and matched WT control cell lines, EpiLC, d2 or d6 PGCLC were dissociated into single-cell solutions with TrypLE and sorted with a Sony SH800Z or a MoFlow high-speed cell sorter (Beckman Coulter) based on appropriate Stella -GFP and Esg1 -tdTomato expression. .. Samples were collected in 150 μl of extraction buffer from the PicoPure RNA isolation kit (Life Technologies) and rapidly frozen on dry ice or liquid nitrogen.

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    Sony sh800z cell sorter
    C8 augments important maturation markers specifically in INS-producing β cells. Quantitative RT-PCR analysis was performed to measure the relative expression levels of the β cell lineage markers INS , UCN3 , and GCK and the transcription factors PDX1 , NKX6.1 , and MAFA from sorted Venus+ and NC cells with or without treatment with C8 and/or bFGF. Briefly, #9–15 hiPSCs were differentiated as described above and treated with 5 μM C8 or 5 μM C8 and 50 ng/mL bFGF in the third stage of differentiation. After differentiation, samples were dissociated with TrypLE and sorted with an <t>SH800Z</t> cell sorter (Sony) for Venus+ and NC cells. Fifty cells from each group were sorted into 5 μL FCP reagent, which is a lysate reagent of the QIAGEN Fast Lane cDNA kit. For cDNA synthesis, 2 μL of the samples were used, with 0.5 μL of the 10 μL cDNA samples used for quantitative RT-PCR. Error bars indicate SD, n = 3. *p
    Sh800z Cell Sorter, supplied by Sony, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sh800z cell sorter/product/Sony
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sh800z cell sorter - by Bioz Stars, 2021-07
    86/100 stars
      Buy from Supplier

    86
    Sony sh800z sorter
    C8 augments important maturation markers specifically in INS-producing β cells. Quantitative RT-PCR analysis was performed to measure the relative expression levels of the β cell lineage markers INS , UCN3 , and GCK and the transcription factors PDX1 , NKX6.1 , and MAFA from sorted Venus+ and NC cells with or without treatment with C8 and/or bFGF. Briefly, #9–15 hiPSCs were differentiated as described above and treated with 5 μM C8 or 5 μM C8 and 50 ng/mL bFGF in the third stage of differentiation. After differentiation, samples were dissociated with TrypLE and sorted with an <t>SH800Z</t> cell sorter (Sony) for Venus+ and NC cells. Fifty cells from each group were sorted into 5 μL FCP reagent, which is a lysate reagent of the QIAGEN Fast Lane cDNA kit. For cDNA synthesis, 2 μL of the samples were used, with 0.5 μL of the 10 μL cDNA samples used for quantitative RT-PCR. Error bars indicate SD, n = 3. *p
    Sh800z Sorter, supplied by Sony, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sh800z sorter/product/Sony
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sh800z sorter - by Bioz Stars, 2021-07
    86/100 stars
      Buy from Supplier

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    C8 augments important maturation markers specifically in INS-producing β cells. Quantitative RT-PCR analysis was performed to measure the relative expression levels of the β cell lineage markers INS , UCN3 , and GCK and the transcription factors PDX1 , NKX6.1 , and MAFA from sorted Venus+ and NC cells with or without treatment with C8 and/or bFGF. Briefly, #9–15 hiPSCs were differentiated as described above and treated with 5 μM C8 or 5 μM C8 and 50 ng/mL bFGF in the third stage of differentiation. After differentiation, samples were dissociated with TrypLE and sorted with an SH800Z cell sorter (Sony) for Venus+ and NC cells. Fifty cells from each group were sorted into 5 μL FCP reagent, which is a lysate reagent of the QIAGEN Fast Lane cDNA kit. For cDNA synthesis, 2 μL of the samples were used, with 0.5 μL of the 10 μL cDNA samples used for quantitative RT-PCR. Error bars indicate SD, n = 3. *p

    Journal: Scientific Reports

    Article Title: An inhibitor of fibroblast growth factor receptor-1 (FGFR1) promotes late-stage terminal differentiation from NGN3+ pancreatic endocrine progenitors

    doi: 10.1038/srep35908

    Figure Lengend Snippet: C8 augments important maturation markers specifically in INS-producing β cells. Quantitative RT-PCR analysis was performed to measure the relative expression levels of the β cell lineage markers INS , UCN3 , and GCK and the transcription factors PDX1 , NKX6.1 , and MAFA from sorted Venus+ and NC cells with or without treatment with C8 and/or bFGF. Briefly, #9–15 hiPSCs were differentiated as described above and treated with 5 μM C8 or 5 μM C8 and 50 ng/mL bFGF in the third stage of differentiation. After differentiation, samples were dissociated with TrypLE and sorted with an SH800Z cell sorter (Sony) for Venus+ and NC cells. Fifty cells from each group were sorted into 5 μL FCP reagent, which is a lysate reagent of the QIAGEN Fast Lane cDNA kit. For cDNA synthesis, 2 μL of the samples were used, with 0.5 μL of the 10 μL cDNA samples used for quantitative RT-PCR. Error bars indicate SD, n = 3. *p

    Article Snippet: The number of Venus- and mCherry-expressing cells was analysed in an SH800Z cell sorter (Sony, Tokyo, Japan).

    Techniques: Quantitative RT-PCR, Expressing