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  • 93
    Addgene inc carboxy terminal 19 amino acids
    Carboxy Terminal 19 Amino Acids, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carboxy terminal 19 amino acids/product/Addgene inc
    Average 93 stars, based on 1 article reviews
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    carboxy terminal 19 amino acids - by Bioz Stars, 2024-06
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    90
    Addgene inc sacas9 transgene
    A, Schematic of gRNA design to target R176Q mutant Ryr2 allele in genomic DNA of R176Q/+ mouse. The mutant codon resulting in the R176Q mutation (red) and the silent Rsr restriction site (green) are marked in the sequence. The guide sequence (22-nt, blue) pairs with the DNA target marked as blue bar upstream of a 5’-NNGRRT adjacent motif (PAM; orange). Cas9 will mediate a double-stranded break ~3 bp upstream of the PAM (red triangle). B, Schematic representation of the <t>SaCas9</t> AAV9 constructs. Original backbone with BbsI cloning sites is used as control.
    Sacas9 Transgene, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sacas9 transgene/product/Addgene inc
    Average 90 stars, based on 1 article reviews
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    sacas9 transgene - by Bioz Stars, 2024-06
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    93
    DSMZ cmv pv 0473
    A, Schematic of gRNA design to target R176Q mutant Ryr2 allele in genomic DNA of R176Q/+ mouse. The mutant codon resulting in the R176Q mutation (red) and the silent Rsr restriction site (green) are marked in the sequence. The guide sequence (22-nt, blue) pairs with the DNA target marked as blue bar upstream of a 5’-NNGRRT adjacent motif (PAM; orange). Cas9 will mediate a double-stranded break ~3 bp upstream of the PAM (red triangle). B, Schematic representation of the <t>SaCas9</t> AAV9 constructs. Original backbone with BbsI cloning sites is used as control.
    Cmv Pv 0473, supplied by DSMZ, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cmv pv 0473/product/DSMZ
    Average 93 stars, based on 1 article reviews
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    cmv pv 0473 - by Bioz Stars, 2024-06
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    90
    DSMZ b trispora dsm 2388
    Strains and plasmids used in this study.
    B Trispora Dsm 2388, supplied by DSMZ, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/b trispora dsm 2388/product/DSMZ
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    b trispora dsm 2388 - by Bioz Stars, 2024-06
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    93
    Sino Biological her2
    Design of FR-engineered e23sFv derivatives. (A) Amino acid sequence alignment of the VL and VH domains of mouse <t>anti-HER2</t> single-chain variable fragment, e23sFv, and their five most homologous counterparts identified in the National Centre for Biotechnology Information protein database. L1-L5 represent VL homologous sequences and H1-H5 represent VH homologous sequences. CDRs and FRs are indicated in columns. The residues that are identical to those of e23sFv are indicated with dashed lines, and missing residues in the CDRs are indicated with asterisks. Non-identical FR residues in e23sFv and all their five homologs in the VL or VH collection are in red. Introduced site-directed mutations are indicated by blue triangles, above which are the corresponding substituted residues. (B) The schematic structure of three e23sFv derivatives. EMEY includes 11 mutated residues in the FRs of e23sFv, as indicated by triangles. EX1 and EX2 represent CDR grafts of e23sFv in the L1-H1 and L2-H2 FR scaffolds, respectively. FR, framework region; VL, light-chain variable region; VH, heavy-chain variable region; CDR, complementarity-determining region.
    Her2, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/her2/product/Sino Biological
    Average 93 stars, based on 1 article reviews
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    her2 - by Bioz Stars, 2024-06
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    Image Search Results


    A, Schematic of gRNA design to target R176Q mutant Ryr2 allele in genomic DNA of R176Q/+ mouse. The mutant codon resulting in the R176Q mutation (red) and the silent Rsr restriction site (green) are marked in the sequence. The guide sequence (22-nt, blue) pairs with the DNA target marked as blue bar upstream of a 5’-NNGRRT adjacent motif (PAM; orange). Cas9 will mediate a double-stranded break ~3 bp upstream of the PAM (red triangle). B, Schematic representation of the SaCas9 AAV9 constructs. Original backbone with BbsI cloning sites is used as control.

    Journal: Circulation research

    Article Title: In Vivo Ryr 2 Editing Corrects Catecholaminergic Polymorphic Ventricular Tachycardia

    doi: 10.1161/CIRCRESAHA.118.313369

    Figure Lengend Snippet: A, Schematic of gRNA design to target R176Q mutant Ryr2 allele in genomic DNA of R176Q/+ mouse. The mutant codon resulting in the R176Q mutation (red) and the silent Rsr restriction site (green) are marked in the sequence. The guide sequence (22-nt, blue) pairs with the DNA target marked as blue bar upstream of a 5’-NNGRRT adjacent motif (PAM; orange). Cas9 will mediate a double-stranded break ~3 bp upstream of the PAM (red triangle). B, Schematic representation of the SaCas9 AAV9 constructs. Original backbone with BbsI cloning sites is used as control.

    Article Snippet: The AAV-CRISPR vector, 1255_pAAV-U6-SA-BbsI-MluI-gRNA-CB-SACas9-HA-OLLAS-spA was generated by the Lagor lab (Addgene plasmid 109320) based on the SaCas9 transgene from px602, Addgene plasmid 61593 (gift of Dr. Feng Zhang).

    Techniques: Mutagenesis, Sequencing, Construct, Clone Assay

    A, Experimental timeline of AAV9 injections and subsequent phenotypical analysis B, Representative examples of simultaneous recording of surface ECG lead I (L-1) and intracardiac ventricular electrogram showing sustained polymorphic ventricular tachycardia in a R176Q/+ mouse treated with placebo (AAV9-Bbsl-SaCas9; left) and normal sinus rhythm in an AAV9-gRNA-SaCas9 treated R176Q/+ mouse (right). C, Bar graph summarizing the incidence of inducible sustained VT in R176Q/+ mice and WT littermates treated with AAV9-Bbsl-SaCas9 or AAV9-gRNA-SaCas9, respectively. Abbreviations: P, postnatal day; SC, subcutaneous. *p<0.05.

    Journal: Circulation research

    Article Title: In Vivo Ryr 2 Editing Corrects Catecholaminergic Polymorphic Ventricular Tachycardia

    doi: 10.1161/CIRCRESAHA.118.313369

    Figure Lengend Snippet: A, Experimental timeline of AAV9 injections and subsequent phenotypical analysis B, Representative examples of simultaneous recording of surface ECG lead I (L-1) and intracardiac ventricular electrogram showing sustained polymorphic ventricular tachycardia in a R176Q/+ mouse treated with placebo (AAV9-Bbsl-SaCas9; left) and normal sinus rhythm in an AAV9-gRNA-SaCas9 treated R176Q/+ mouse (right). C, Bar graph summarizing the incidence of inducible sustained VT in R176Q/+ mice and WT littermates treated with AAV9-Bbsl-SaCas9 or AAV9-gRNA-SaCas9, respectively. Abbreviations: P, postnatal day; SC, subcutaneous. *p<0.05.

    Article Snippet: The AAV-CRISPR vector, 1255_pAAV-U6-SA-BbsI-MluI-gRNA-CB-SACas9-HA-OLLAS-spA was generated by the Lagor lab (Addgene plasmid 109320) based on the SaCas9 transgene from px602, Addgene plasmid 61593 (gift of Dr. Feng Zhang).

    Techniques:

    A, Diagram showing where PCR primers bind to sequences in exons 3-6 upstream of mutation R176Q in exon 8 (green) and primers targeting exons 11-13 downstream of R176Q (orange). B, Levels of RyR2 mRNA in cardiac tissue from R176Q/+ mice or WT littermates, treated with BbsI-SaCas9 or gRNA-SaCas9. C, Western blot analysis of RyR2 and JPH2 protein levels in heart tissue from R176Q/+ mice or WT littermates. D, Corresponding quantifications of RyR2, and E, JPH2 proteins levels normalized to GAPDH. *p<0.05.

    Journal: Circulation research

    Article Title: In Vivo Ryr 2 Editing Corrects Catecholaminergic Polymorphic Ventricular Tachycardia

    doi: 10.1161/CIRCRESAHA.118.313369

    Figure Lengend Snippet: A, Diagram showing where PCR primers bind to sequences in exons 3-6 upstream of mutation R176Q in exon 8 (green) and primers targeting exons 11-13 downstream of R176Q (orange). B, Levels of RyR2 mRNA in cardiac tissue from R176Q/+ mice or WT littermates, treated with BbsI-SaCas9 or gRNA-SaCas9. C, Western blot analysis of RyR2 and JPH2 protein levels in heart tissue from R176Q/+ mice or WT littermates. D, Corresponding quantifications of RyR2, and E, JPH2 proteins levels normalized to GAPDH. *p<0.05.

    Article Snippet: The AAV-CRISPR vector, 1255_pAAV-U6-SA-BbsI-MluI-gRNA-CB-SACas9-HA-OLLAS-spA was generated by the Lagor lab (Addgene plasmid 109320) based on the SaCas9 transgene from px602, Addgene plasmid 61593 (gift of Dr. Feng Zhang).

    Techniques: Mutagenesis, Western Blot

    A, Schematic picture showing gRNA-SaCas9 and “reporter” AAV9 co-injected in R176Q/+ mice. The “reporter” virus with cloned targeted sequence was cut by gRNA-SaCas9 but not the control BbsI-SaCas9. The mCherry protein is only expressed in cardiomycytes with SaCas9 expression. B, Representative confocal images of ventricle myocytes from R176Q/+ mice co-injected with Bbsl-SaCas9 (control) + “reporter” and gRNA-SaCas9 (editing) + “reporter”, merged with bright field (BF) and red fluorescence (RFP) signal overlay. Scale bar, 100 μm. C, Bar graph showing percentages of mCherry-positive cardiomyocytes. D-F, Summary data for Ca2+ spark frequency, SR Ca2+ load, and SR Ca2+ transient amplitude. *p<0.05, **p<0.01, ***p<0.001.

    Journal: Circulation research

    Article Title: In Vivo Ryr 2 Editing Corrects Catecholaminergic Polymorphic Ventricular Tachycardia

    doi: 10.1161/CIRCRESAHA.118.313369

    Figure Lengend Snippet: A, Schematic picture showing gRNA-SaCas9 and “reporter” AAV9 co-injected in R176Q/+ mice. The “reporter” virus with cloned targeted sequence was cut by gRNA-SaCas9 but not the control BbsI-SaCas9. The mCherry protein is only expressed in cardiomycytes with SaCas9 expression. B, Representative confocal images of ventricle myocytes from R176Q/+ mice co-injected with Bbsl-SaCas9 (control) + “reporter” and gRNA-SaCas9 (editing) + “reporter”, merged with bright field (BF) and red fluorescence (RFP) signal overlay. Scale bar, 100 μm. C, Bar graph showing percentages of mCherry-positive cardiomyocytes. D-F, Summary data for Ca2+ spark frequency, SR Ca2+ load, and SR Ca2+ transient amplitude. *p<0.05, **p<0.01, ***p<0.001.

    Article Snippet: The AAV-CRISPR vector, 1255_pAAV-U6-SA-BbsI-MluI-gRNA-CB-SACas9-HA-OLLAS-spA was generated by the Lagor lab (Addgene plasmid 109320) based on the SaCas9 transgene from px602, Addgene plasmid 61593 (gift of Dr. Feng Zhang).

    Techniques: Injection, Clone Assay, Sequencing, Expressing, Fluorescence

    Strains and plasmids used in this study.

    Journal: Scientific Reports

    Article Title: Manipulation of the precursor supply for high-level production of longifolene by metabolically engineered Escherichia coli

    doi: 10.1038/s41598-018-36495-w

    Figure Lengend Snippet: Strains and plasmids used in this study.

    Article Snippet: B. trispora DSM-2388 , Mating type - , German Collection of Microorganisms and Cell Cultures (DSMZ).

    Techniques:

    Design of FR-engineered e23sFv derivatives. (A) Amino acid sequence alignment of the VL and VH domains of mouse anti-HER2 single-chain variable fragment, e23sFv, and their five most homologous counterparts identified in the National Centre for Biotechnology Information protein database. L1-L5 represent VL homologous sequences and H1-H5 represent VH homologous sequences. CDRs and FRs are indicated in columns. The residues that are identical to those of e23sFv are indicated with dashed lines, and missing residues in the CDRs are indicated with asterisks. Non-identical FR residues in e23sFv and all their five homologs in the VL or VH collection are in red. Introduced site-directed mutations are indicated by blue triangles, above which are the corresponding substituted residues. (B) The schematic structure of three e23sFv derivatives. EMEY includes 11 mutated residues in the FRs of e23sFv, as indicated by triangles. EX1 and EX2 represent CDR grafts of e23sFv in the L1-H1 and L2-H2 FR scaffolds, respectively. FR, framework region; VL, light-chain variable region; VH, heavy-chain variable region; CDR, complementarity-determining region.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Syngeneic homograft of framework regions enhances the affinity of the mouse anti-human epidermal receptor 2 single-chain antibody e23sFv

    doi: 10.3892/etm.2020.9568

    Figure Lengend Snippet: Design of FR-engineered e23sFv derivatives. (A) Amino acid sequence alignment of the VL and VH domains of mouse anti-HER2 single-chain variable fragment, e23sFv, and their five most homologous counterparts identified in the National Centre for Biotechnology Information protein database. L1-L5 represent VL homologous sequences and H1-H5 represent VH homologous sequences. CDRs and FRs are indicated in columns. The residues that are identical to those of e23sFv are indicated with dashed lines, and missing residues in the CDRs are indicated with asterisks. Non-identical FR residues in e23sFv and all their five homologs in the VL or VH collection are in red. Introduced site-directed mutations are indicated by blue triangles, above which are the corresponding substituted residues. (B) The schematic structure of three e23sFv derivatives. EMEY includes 11 mutated residues in the FRs of e23sFv, as indicated by triangles. EX1 and EX2 represent CDR grafts of e23sFv in the L1-H1 and L2-H2 FR scaffolds, respectively. FR, framework region; VL, light-chain variable region; VH, heavy-chain variable region; CDR, complementarity-determining region.

    Article Snippet: Recombinant human HER2 (Sino Biological; 500 ng per well) was immobilized on 96-well plates at 4˚C for 16 h. The HER2-coated plates were blocked with 1% BSA (Sigma-Aldrich; Merck KGaA) at room temperature for 1 h and incubated with the three-fold-serially diluted e23sFv derivatives from 3 µM for 4 h at room temperature.

    Techniques: Sequencing

    In vitro binding of the e23sFv-derived scFvs to recombinant HER2. (A) Affinity measurement by ELISA. HER2-coated microplates were incubated with the e23sFv derivatives at various concentrations, and the bound scFvs were detected using an anti-His antibody. scFv15 served as the negative control. (B) One-shot kinetics of SPR. Five sensorgrams indicated the response of HER2-immobilized sensor chips with five diluted concentrations of the e23sFv derivatives. (C) Comparison of K on , K off and K D of the e23sFv derivatives calculated from SPR sensing. scFv, single-chain variable fragment; SPR, surface plasmon resonance; RU, resonance unit; K on , association rate constant; K off , dissociation rate constant, K D , equilibrium constant; Chi 2 , goodness-of-fit between the binding model and theoretical affinity; OD, optical density.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Syngeneic homograft of framework regions enhances the affinity of the mouse anti-human epidermal receptor 2 single-chain antibody e23sFv

    doi: 10.3892/etm.2020.9568

    Figure Lengend Snippet: In vitro binding of the e23sFv-derived scFvs to recombinant HER2. (A) Affinity measurement by ELISA. HER2-coated microplates were incubated with the e23sFv derivatives at various concentrations, and the bound scFvs were detected using an anti-His antibody. scFv15 served as the negative control. (B) One-shot kinetics of SPR. Five sensorgrams indicated the response of HER2-immobilized sensor chips with five diluted concentrations of the e23sFv derivatives. (C) Comparison of K on , K off and K D of the e23sFv derivatives calculated from SPR sensing. scFv, single-chain variable fragment; SPR, surface plasmon resonance; RU, resonance unit; K on , association rate constant; K off , dissociation rate constant, K D , equilibrium constant; Chi 2 , goodness-of-fit between the binding model and theoretical affinity; OD, optical density.

    Article Snippet: Recombinant human HER2 (Sino Biological; 500 ng per well) was immobilized on 96-well plates at 4˚C for 16 h. The HER2-coated plates were blocked with 1% BSA (Sigma-Aldrich; Merck KGaA) at room temperature for 1 h and incubated with the three-fold-serially diluted e23sFv derivatives from 3 µM for 4 h at room temperature.

    Techniques: In Vitro, Binding Assay, Derivative Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Incubation, Negative Control, SPR Assay

    Binding of the e23sFv-derived single-chain variable fragments to HER2 on the cell surface. HER2-positive cells (BT-474 and SKOV-3 cells) and HER2-negative cells (MCF-7 cells) were incubated with FITC-labelled e23sFv derivatives and subjected to flow cytometry analysis. (A) Representative dataset of one-parameter histograms. For the isotype control, FITC-labelled scFv15 against HBsAg was used. For the positive control, a commercial FITC-conjugated anti-HER2 antibody was used. (B) Statistical analysis from three independent parallel experiments. * P<0.05; ** P<0.01 and *** P<0.001. ns, non-significant.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Syngeneic homograft of framework regions enhances the affinity of the mouse anti-human epidermal receptor 2 single-chain antibody e23sFv

    doi: 10.3892/etm.2020.9568

    Figure Lengend Snippet: Binding of the e23sFv-derived single-chain variable fragments to HER2 on the cell surface. HER2-positive cells (BT-474 and SKOV-3 cells) and HER2-negative cells (MCF-7 cells) were incubated with FITC-labelled e23sFv derivatives and subjected to flow cytometry analysis. (A) Representative dataset of one-parameter histograms. For the isotype control, FITC-labelled scFv15 against HBsAg was used. For the positive control, a commercial FITC-conjugated anti-HER2 antibody was used. (B) Statistical analysis from three independent parallel experiments. * P<0.05; ** P<0.01 and *** P<0.001. ns, non-significant.

    Article Snippet: Recombinant human HER2 (Sino Biological; 500 ng per well) was immobilized on 96-well plates at 4˚C for 16 h. The HER2-coated plates were blocked with 1% BSA (Sigma-Aldrich; Merck KGaA) at room temperature for 1 h and incubated with the three-fold-serially diluted e23sFv derivatives from 3 µM for 4 h at room temperature.

    Techniques: Binding Assay, Derivative Assay, Incubation, Flow Cytometry, Positive Control

    Internalization of the e23sFv-derived single-chain variable fragments by HER2-positive cells. Following incubation with FITC-labelled e23sFv derivatives, BT-474, SKOV-3 and MCF-7 cells were observed under fluorescence microscopy. MCF-7 cells served as the HER2-negative cell controls and scFv15 was the non-specific binding control. Scale bar, 100 µm. The data are representative of at least three independent experiments.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Syngeneic homograft of framework regions enhances the affinity of the mouse anti-human epidermal receptor 2 single-chain antibody e23sFv

    doi: 10.3892/etm.2020.9568

    Figure Lengend Snippet: Internalization of the e23sFv-derived single-chain variable fragments by HER2-positive cells. Following incubation with FITC-labelled e23sFv derivatives, BT-474, SKOV-3 and MCF-7 cells were observed under fluorescence microscopy. MCF-7 cells served as the HER2-negative cell controls and scFv15 was the non-specific binding control. Scale bar, 100 µm. The data are representative of at least three independent experiments.

    Article Snippet: Recombinant human HER2 (Sino Biological; 500 ng per well) was immobilized on 96-well plates at 4˚C for 16 h. The HER2-coated plates were blocked with 1% BSA (Sigma-Aldrich; Merck KGaA) at room temperature for 1 h and incubated with the three-fold-serially diluted e23sFv derivatives from 3 µM for 4 h at room temperature.

    Techniques: Derivative Assay, Incubation, Fluorescence, Microscopy, Binding Assay

    Docking mechanism of the enhanced EX1-HER2 interaction. (A-D) In silico docking of e23sFv, EMEY, EX1 and EX2 and their interactions with the predicted surface models of the HER2 ECD. All the scFv fragments form distinct but overlapping interfaces with domain IV of the HER2 ECD. The 3D structures of (A) e23sFv, (B) EMEY, (C) EX1 and (D) EX2 are presented as coloured ribbons. (E) Binding energy with HER2 and the predicted binding epitopes of all the scFv fragments. ECD, extracellular domain; scFv, single-chain variable fragment.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Syngeneic homograft of framework regions enhances the affinity of the mouse anti-human epidermal receptor 2 single-chain antibody e23sFv

    doi: 10.3892/etm.2020.9568

    Figure Lengend Snippet: Docking mechanism of the enhanced EX1-HER2 interaction. (A-D) In silico docking of e23sFv, EMEY, EX1 and EX2 and their interactions with the predicted surface models of the HER2 ECD. All the scFv fragments form distinct but overlapping interfaces with domain IV of the HER2 ECD. The 3D structures of (A) e23sFv, (B) EMEY, (C) EX1 and (D) EX2 are presented as coloured ribbons. (E) Binding energy with HER2 and the predicted binding epitopes of all the scFv fragments. ECD, extracellular domain; scFv, single-chain variable fragment.

    Article Snippet: Recombinant human HER2 (Sino Biological; 500 ng per well) was immobilized on 96-well plates at 4˚C for 16 h. The HER2-coated plates were blocked with 1% BSA (Sigma-Aldrich; Merck KGaA) at room temperature for 1 h and incubated with the three-fold-serially diluted e23sFv derivatives from 3 µM for 4 h at room temperature.

    Techniques: In Silico, Binding Assay