programmable digital attenuator operating Search Results


anti nanog antibody  (Developmental Studies Hybridoma Bank)
93
Developmental Studies Hybridoma Bank anti nanog antibody
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
Anti Nanog Antibody, supplied by Developmental Studies Hybridoma Bank, 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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programmable digital attenuator operating  (Mini-Circuits)
94
Mini-Circuits programmable digital attenuator operating
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
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affinity attenuated omci 9  (CompTech Computer Technologies)
90
CompTech Computer Technologies affinity attenuated omci 9
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
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computer adaptive scales  (ProMIS Neurosciences)
90
ProMIS Neurosciences computer adaptive scales
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
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programmable attenuator  (Tucker-Davis Tech)
90
Tucker-Davis Tech programmable attenuator
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
Programmable Attenuator, supplied by Tucker-Davis Tech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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one attension theta optical tensiometer  (Biolin Scientific)
90
Biolin Scientific one attension theta optical tensiometer
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
One Attension Theta Optical Tensiometer, supplied by Biolin Scientific, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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information communication (icts)  (Thermo Fisher)
90
Thermo Fisher information communication (icts)
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
Information Communication (Icts), supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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one-reflection attenuated total reflection computer-controlled bruker spectrometer  (Bruker Corporation)
90
Bruker Corporation one-reflection attenuated total reflection computer-controlled bruker spectrometer
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
One Reflection Attenuated Total Reflection Computer Controlled Bruker Spectrometer, supplied by Bruker Corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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computer algebra systems (cas)  (MathWorks Inc)
90
MathWorks Inc computer algebra systems (cas)
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
Computer Algebra Systems (Cas), supplied by MathWorks Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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digital program for attention-deficit/hyperactivity disorder (adhd)  (Akili Interactive Labs)
90
Akili Interactive Labs digital program for attention-deficit/hyperactivity disorder (adhd)
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
Digital Program For Attention Deficit/Hyperactivity Disorder (Adhd), supplied by Akili Interactive Labs, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human c5 protein  (CompTech Computer Technologies)
90
CompTech Computer Technologies human c5 protein
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
Human C5 Protein, supplied by CompTech Computer Technologies, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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eprime 1.1  (psychology software tools)
90
psychology software tools eprime 1.1
( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, <t>green),</t> <t>OCT3/4</t> (center, yellow), and <t>NANOG</t> (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.
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Image Search Results


( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, green), OCT3/4 (center, yellow), and NANOG (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.

Journal: eLife

Article Title: Endogenous tagging using split mNeonGreen in human iPSCs for live imaging studies

doi: 10.7554/eLife.92819

Figure Lengend Snippet: ( A ) A schematic representation of the split tagging strategy used in this study. An expression cassette carrying the mNG2 1-10 fragment was integrated at the AAVS1 locus in 201B7 iPSCs to generate the parental split mNeonGreen2 cell line (smNG2-P). Proteins of interest were tagged endogenously with the mNG2 11 fragment to visualize their expression and localization. The structure of mNeonGreen was generated from PDB (Protein Data Bank, structure identifier 5 LTP; ). ( B ) A schematic of the mNG2 1-10 expression cassette is shown with the CAG promoter (blue) and sequences for stable expression (dark gray), Puromycin resistance marker (pink), and homology arms (light gray) for integration at the AAVS1 locus. Abbreviations: SA = splicing acceptor, T2A=Thosea asigna virus 2A peptide, bGH = bovine growth hormone poly-adenylation signal, β-glo=rabbit β-globin poly-adenylation signal. ( C ) Sequencing chromatograms show the edited AAVS1 allele junctions in the smNG2-P cell line (bottom) compared to the WT allele (top). ( D ) Representative G-banding karyotype of the smNG2-P cell line shows that the edited cells have a normal karyotype (46, XX). ( E ) Flow cytometry plots show smNG2-P cells stained for pluripotency markers TRA-1–60 (left, green), OCT3/4 (center, yellow), and NANOG (right, red) or a secondary antibody control (blue). ( F ) Flow cytometry plots show differentiated smNG2-P cells stained for PAX6 (ectoderm; left, green), NCAM (mesoderm; center, yellow), and FOXA2 (endoderm; right, red) compared to undifferentiated cells (blue) and unstained controls (dark colors). ( G ) Fluorescent and brightfield images show fluorescence complementation after transfecting split mNeonGreen2 parental cell line (smNG2-P) cells with a plasmid expressing H2B fused to mNG2 11 . The scale bar is 10 microns.

Article Snippet: The following antibody dilutions were prepared in permeabilization/blocking solution or blocking solution: 1:25 anti-NANOG antibody (1 μg/mL final concentration, PCRP-NANOGP1-2D8, DSHB), 1:20 anti-OCT3/4 antibody (10 μg/mL final concentration; Santa Cruz Biotechnology), or 1:20 Alexa488-conjugated anti-TRA-1–60 antibody (7.5 μg/mL final concentration; STEMCELL Technologies).

Techniques: Expressing, Generated, Marker, Virus, Sequencing, Flow Cytometry, Staining, Control, Fluorescence, Plasmid Preparation

Journal: eLife

Article Title: Endogenous tagging using split mNeonGreen in human iPSCs for live imaging studies

doi: 10.7554/eLife.92819

Figure Lengend Snippet:

Article Snippet: The following antibody dilutions were prepared in permeabilization/blocking solution or blocking solution: 1:25 anti-NANOG antibody (1 μg/mL final concentration, PCRP-NANOGP1-2D8, DSHB), 1:20 anti-OCT3/4 antibody (10 μg/mL final concentration; Santa Cruz Biotechnology), or 1:20 Alexa488-conjugated anti-TRA-1–60 antibody (7.5 μg/mL final concentration; STEMCELL Technologies).

Techniques: Recombinant, Sequencing, Ligation, Digital PCR, Software, Polymer, Imaging