dynabeads m 270 streptavidin coated magnetic beads Thermo Fisher Search Results


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    Thermo Fisher streptavidin coated magnetic bead
    Transverse magnetic tweezers setup and Dps mediated DNA condensation. ( a ) Figure shows an illustration of single-DNA stretching experiment using magnetic tweezers. One biotinylated DNA end is attached to a <t>streptavidin</t> functionalised coverslip and the other biotinylated DNA end attached to a streptavidin coated paramagnetic bead. In the shaded area of 2 micrometers from the coverslip edge, bead image cannot be obtained. Force is exerted by using a permanent magnet and force is adjusted by moving the position of magnet. At different forces, the corresponding extension of DNA is recorded. ( b ) Force-extension curves obtained on a 48,502 bp λ-DNA during force-decrease (solid symbols) and subsequent force-increase scan (open symbols) before and after incubated with 50 nM Dps, 500 nM Dps and 5000 nM Dps in 50 mM KCl, pH 7.5 at 23 °C. Black data shows force extension curve of naked DNA as control where force-decreased and force increased curves overlapped. The black solid curve is a fitting curve by the Worm-like chain DNA polymer model with a persistence length of 50 nm using the Marko-Siggia formula 35 36 . Force extension curves after protein incubation are plotted with coloured symbols, coloured lines are connecting lines between each data points for better presentation. Data obtained from three independent experiments at each concentration are shown in the same colour indicated by solid, dashed, and dotted connecting lines. The non-overlapping force-decreased and force increased curves shows that force-extension curves of the DNA interacting with Dps do not reach equilibrium at our force-scanning experimental time scale.
    Streptavidin Coated Magnetic Bead, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin coated magnetic bead/product/Thermo Fisher
    Average 90 stars, based on 39 article reviews
    Price from $9.99 to $1999.99
    streptavidin coated magnetic bead - by Bioz Stars, 2020-02
    90/100 stars
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    83
    Thermo Fisher ap model streptavidin coated magnetic beads
    Development of a classical pathway C3 and C5 convertase model. (A) In the CP convertase model, <t>streptavidin</t> beads are labeled with biotinylated DNP which serves as a model antigen. Addition of IgG1 recognizing DNP and complement proteins results in formation of C4b2a and C4b2a(C3b) n convertases on the bead surface, which convert C3 and C5. (B) Only in the presence of all CP components (antigen, IgG, and CP proteins) C3 and C5 are converted, as measured by calcium mobilization in U937-C3aR and U937-C5aR cells, respectively. (C) C3 conversion via CP convertases results in release of C3a in the supernatant (as shown by calcium mobilization) and (D) C3b deposition on the bead surface (as shown by flow cytometry). Identical amounts of the non-reactive C3 variants C3b H2O and C3 MA do not bind to the bead surface. (E) C5 convertase activity increases with the level of deposited C3b molecules on the beads surface, but is not affected by C3b H2O or C3 MA in solution. Uncoupling C3 and C5 conversion by introduction of an extra washing step does not alter C5 conversion, indicating that CP C5 conversion only depends on deposited C3b molecules around existing C3 convertases. (B–E) Data of three independent experiments, presented as mean ± SD.
    Ap Model Streptavidin Coated Magnetic Beads, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 83/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ap model streptavidin coated magnetic beads/product/Thermo Fisher
    Average 83 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    ap model streptavidin coated magnetic beads - by Bioz Stars, 2020-02
    83/100 stars
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    81
    Thermo Fisher c3b coated beads streptavidin coated magnetic beads
    A novel bead-based assay model for purified alternative pathway (AP) C5 convertases. a Proposed model for assembly of C5 convertases in the AP. Surface-bound C3 convertase (C3bBb) cleaves multiple C3 molecules into <t>C3b</t> that covalently binds to target surfaces via the reactive thioester (red dot). Association of deposited C3b molecules with the existing C3 convertase gives rise to multimeric complexes (C3b-C3b n ) that, together with Bb, can convert C5. The precise arrangement of surface-specific C5 convertases is currently unknown. In the novel C5 convertase assay model described in this study, C3b molecules are site-specifically biotinylated via the thioester and loaded on bacteria-sized <t>streptavidin</t> beads (2.8 μm) to mimic their natural density and orientation on target surfaces. b Loading of streptavidin beads with biotinylated C3b was analyzed by flow cytometry or immunoblotting (below). c C5 convertase activity of C3b-coated beads that were incubated with factor B (FB), factor D (FD) (together needed to form Bb) and C5. Conversion of C5 was determined by measuring release of C5a in the supernatant using a calcium mobilization assay with U937-C5aR cells. Values represent absolute C5a flux (mean fluorescence of stimulated cells subtracted by the mean fluorescence before stimulus). d C5 convertase activity of C3b molecules on beads versus C3b molecules in solution. The amount of C3b molecules in solution was adjusted to the levels of C3b loaded onto the beads (relative C3b-biotin levels) and both were incubated with FB, FD and C5. b – d Data of three independent experiments, presented as means ± standard deviation (SD). Immunoblot is a representative of three independent experiments
    C3b Coated Beads Streptavidin Coated Magnetic Beads, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 81/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/c3b coated beads streptavidin coated magnetic beads/product/Thermo Fisher
    Average 81 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    c3b coated beads streptavidin coated magnetic beads - by Bioz Stars, 2020-02
    81/100 stars
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    Image Search Results


    Transverse magnetic tweezers setup and Dps mediated DNA condensation. ( a ) Figure shows an illustration of single-DNA stretching experiment using magnetic tweezers. One biotinylated DNA end is attached to a streptavidin functionalised coverslip and the other biotinylated DNA end attached to a streptavidin coated paramagnetic bead. In the shaded area of 2 micrometers from the coverslip edge, bead image cannot be obtained. Force is exerted by using a permanent magnet and force is adjusted by moving the position of magnet. At different forces, the corresponding extension of DNA is recorded. ( b ) Force-extension curves obtained on a 48,502 bp λ-DNA during force-decrease (solid symbols) and subsequent force-increase scan (open symbols) before and after incubated with 50 nM Dps, 500 nM Dps and 5000 nM Dps in 50 mM KCl, pH 7.5 at 23 °C. Black data shows force extension curve of naked DNA as control where force-decreased and force increased curves overlapped. The black solid curve is a fitting curve by the Worm-like chain DNA polymer model with a persistence length of 50 nm using the Marko-Siggia formula 35 36 . Force extension curves after protein incubation are plotted with coloured symbols, coloured lines are connecting lines between each data points for better presentation. Data obtained from three independent experiments at each concentration are shown in the same colour indicated by solid, dashed, and dotted connecting lines. The non-overlapping force-decreased and force increased curves shows that force-extension curves of the DNA interacting with Dps do not reach equilibrium at our force-scanning experimental time scale.

    Journal: Scientific Reports

    Article Title: Regulation of Bacterial DNA Packaging in Early Stationary Phase by Competitive DNA Binding of Dps and IHF

    doi: 10.1038/srep18146

    Figure Lengend Snippet: Transverse magnetic tweezers setup and Dps mediated DNA condensation. ( a ) Figure shows an illustration of single-DNA stretching experiment using magnetic tweezers. One biotinylated DNA end is attached to a streptavidin functionalised coverslip and the other biotinylated DNA end attached to a streptavidin coated paramagnetic bead. In the shaded area of 2 micrometers from the coverslip edge, bead image cannot be obtained. Force is exerted by using a permanent magnet and force is adjusted by moving the position of magnet. At different forces, the corresponding extension of DNA is recorded. ( b ) Force-extension curves obtained on a 48,502 bp λ-DNA during force-decrease (solid symbols) and subsequent force-increase scan (open symbols) before and after incubated with 50 nM Dps, 500 nM Dps and 5000 nM Dps in 50 mM KCl, pH 7.5 at 23 °C. Black data shows force extension curve of naked DNA as control where force-decreased and force increased curves overlapped. The black solid curve is a fitting curve by the Worm-like chain DNA polymer model with a persistence length of 50 nm using the Marko-Siggia formula 35 36 . Force extension curves after protein incubation are plotted with coloured symbols, coloured lines are connecting lines between each data points for better presentation. Data obtained from three independent experiments at each concentration are shown in the same colour indicated by solid, dashed, and dotted connecting lines. The non-overlapping force-decreased and force increased curves shows that force-extension curves of the DNA interacting with Dps do not reach equilibrium at our force-scanning experimental time scale.

    Article Snippet: One end of the biotinylated DNA was tethered to a streptavidin functionalized surface, and the other end to a streptavidin-coated magnetic bead (Dynabeads® M-270 Streptavidin, Invitrogen). shows the schematic diagram of the magnetic tweezers setup.

    Techniques: Incubation, Concentration Assay

    Development of a classical pathway C3 and C5 convertase model. (A) In the CP convertase model, streptavidin beads are labeled with biotinylated DNP which serves as a model antigen. Addition of IgG1 recognizing DNP and complement proteins results in formation of C4b2a and C4b2a(C3b) n convertases on the bead surface, which convert C3 and C5. (B) Only in the presence of all CP components (antigen, IgG, and CP proteins) C3 and C5 are converted, as measured by calcium mobilization in U937-C3aR and U937-C5aR cells, respectively. (C) C3 conversion via CP convertases results in release of C3a in the supernatant (as shown by calcium mobilization) and (D) C3b deposition on the bead surface (as shown by flow cytometry). Identical amounts of the non-reactive C3 variants C3b H2O and C3 MA do not bind to the bead surface. (E) C5 convertase activity increases with the level of deposited C3b molecules on the beads surface, but is not affected by C3b H2O or C3 MA in solution. Uncoupling C3 and C5 conversion by introduction of an extra washing step does not alter C5 conversion, indicating that CP C5 conversion only depends on deposited C3b molecules around existing C3 convertases. (B–E) Data of three independent experiments, presented as mean ± SD.

    Journal: Frontiers in Immunology

    Article Title: Functional Characterization of Alternative and Classical Pathway C3/C5 Convertase Activity and Inhibition Using Purified Models

    doi: 10.3389/fimmu.2018.01691

    Figure Lengend Snippet: Development of a classical pathway C3 and C5 convertase model. (A) In the CP convertase model, streptavidin beads are labeled with biotinylated DNP which serves as a model antigen. Addition of IgG1 recognizing DNP and complement proteins results in formation of C4b2a and C4b2a(C3b) n convertases on the bead surface, which convert C3 and C5. (B) Only in the presence of all CP components (antigen, IgG, and CP proteins) C3 and C5 are converted, as measured by calcium mobilization in U937-C3aR and U937-C5aR cells, respectively. (C) C3 conversion via CP convertases results in release of C3a in the supernatant (as shown by calcium mobilization) and (D) C3b deposition on the bead surface (as shown by flow cytometry). Identical amounts of the non-reactive C3 variants C3b H2O and C3 MA do not bind to the bead surface. (E) C5 convertase activity increases with the level of deposited C3b molecules on the beads surface, but is not affected by C3b H2O or C3 MA in solution. Uncoupling C3 and C5 conversion by introduction of an extra washing step does not alter C5 conversion, indicating that CP C5 conversion only depends on deposited C3b molecules around existing C3 convertases. (B–E) Data of three independent experiments, presented as mean ± SD.

    Article Snippet: C3 and C5 Conversion in AP Model Streptavidin-coated magnetic beads (Dynabeads M-270 Streptavidin, Invitrogen) were washed once in VBS-T/Mg [Veronal Buffered Saline pH 7.4, 2.5 mM MgCl2 , 0.05% (v/v) Tween].

    Techniques: Labeling, Flow Cytometry, Cytometry, Activity Assay

    A novel bead-based assay model for purified alternative pathway (AP) C5 convertases. a Proposed model for assembly of C5 convertases in the AP. Surface-bound C3 convertase (C3bBb) cleaves multiple C3 molecules into C3b that covalently binds to target surfaces via the reactive thioester (red dot). Association of deposited C3b molecules with the existing C3 convertase gives rise to multimeric complexes (C3b-C3b n ) that, together with Bb, can convert C5. The precise arrangement of surface-specific C5 convertases is currently unknown. In the novel C5 convertase assay model described in this study, C3b molecules are site-specifically biotinylated via the thioester and loaded on bacteria-sized streptavidin beads (2.8 μm) to mimic their natural density and orientation on target surfaces. b Loading of streptavidin beads with biotinylated C3b was analyzed by flow cytometry or immunoblotting (below). c C5 convertase activity of C3b-coated beads that were incubated with factor B (FB), factor D (FD) (together needed to form Bb) and C5. Conversion of C5 was determined by measuring release of C5a in the supernatant using a calcium mobilization assay with U937-C5aR cells. Values represent absolute C5a flux (mean fluorescence of stimulated cells subtracted by the mean fluorescence before stimulus). d C5 convertase activity of C3b molecules on beads versus C3b molecules in solution. The amount of C3b molecules in solution was adjusted to the levels of C3b loaded onto the beads (relative C3b-biotin levels) and both were incubated with FB, FD and C5. b – d Data of three independent experiments, presented as means ± standard deviation (SD). Immunoblot is a representative of three independent experiments

    Journal: BMC Biology

    Article Title: Molecular insights into the surface-specific arrangement of complement C5 convertase enzymes

    doi: 10.1186/s12915-015-0203-8

    Figure Lengend Snippet: A novel bead-based assay model for purified alternative pathway (AP) C5 convertases. a Proposed model for assembly of C5 convertases in the AP. Surface-bound C3 convertase (C3bBb) cleaves multiple C3 molecules into C3b that covalently binds to target surfaces via the reactive thioester (red dot). Association of deposited C3b molecules with the existing C3 convertase gives rise to multimeric complexes (C3b-C3b n ) that, together with Bb, can convert C5. The precise arrangement of surface-specific C5 convertases is currently unknown. In the novel C5 convertase assay model described in this study, C3b molecules are site-specifically biotinylated via the thioester and loaded on bacteria-sized streptavidin beads (2.8 μm) to mimic their natural density and orientation on target surfaces. b Loading of streptavidin beads with biotinylated C3b was analyzed by flow cytometry or immunoblotting (below). c C5 convertase activity of C3b-coated beads that were incubated with factor B (FB), factor D (FD) (together needed to form Bb) and C5. Conversion of C5 was determined by measuring release of C5a in the supernatant using a calcium mobilization assay with U937-C5aR cells. Values represent absolute C5a flux (mean fluorescence of stimulated cells subtracted by the mean fluorescence before stimulus). d C5 convertase activity of C3b molecules on beads versus C3b molecules in solution. The amount of C3b molecules in solution was adjusted to the levels of C3b loaded onto the beads (relative C3b-biotin levels) and both were incubated with FB, FD and C5. b – d Data of three independent experiments, presented as means ± standard deviation (SD). Immunoblot is a representative of three independent experiments

    Article Snippet: Preparation of C3b-coated beads Streptavidin-coated magnetic beads (Dynabeads M-270 Streptavidin, Invitrogen (Carlsbad, CA, USA), 2.8 μm diameter) were washed twice and suspended in VBS-T+ (veronal buffered saline: 2 mM veronal, 145 mM NaCl, pH 7.4 (VBS, pH 7.4) containing 2.5 mM MgCl2 and 0.05 % Tween).

    Techniques: Bead-based Assay, Purification, Convertase Assay, Flow Cytometry, Cytometry, Activity Assay, Incubation, Calcium Mobilization Assay, Fluorescence, Standard Deviation

    C5 binds to C3b-coated beads. C3b-coated streptavidin beads were incubated with C5 (in the absence of FB and FD). Binding of C5 to C3b beads was determined by a Western blotting or b flow cytometry. a is a representative gel of three independent experiments; b shows data of three independent experiments, presented as means ± standard deviation (SD)

    Journal: BMC Biology

    Article Title: Molecular insights into the surface-specific arrangement of complement C5 convertase enzymes

    doi: 10.1186/s12915-015-0203-8

    Figure Lengend Snippet: C5 binds to C3b-coated beads. C3b-coated streptavidin beads were incubated with C5 (in the absence of FB and FD). Binding of C5 to C3b beads was determined by a Western blotting or b flow cytometry. a is a representative gel of three independent experiments; b shows data of three independent experiments, presented as means ± standard deviation (SD)

    Article Snippet: Preparation of C3b-coated beads Streptavidin-coated magnetic beads (Dynabeads M-270 Streptavidin, Invitrogen (Carlsbad, CA, USA), 2.8 μm diameter) were washed twice and suspended in VBS-T+ (veronal buffered saline: 2 mM veronal, 145 mM NaCl, pH 7.4 (VBS, pH 7.4) containing 2.5 mM MgCl2 and 0.05 % Tween).

    Techniques: Incubation, Binding Assay, Western Blot, Flow Cytometry, Cytometry, Standard Deviation

    High surface density of C3b is critical for C5 convertase activity. a Preparation of beads with different densities of C3b. b Mixing a fixed amount of C3b molecules with increasing numbers of streptavidin beads results in lower C3b densities per bead (top, flow cytometry) while total levels of C3b per sample are equal (below, immunoblot). c C5 convertase activity on streptavidin beads with different C3b densities. d C5 convertase activity plotted against the absolute number of C3b molecules per μm 2 (calculated from the results in c ). b – d Data of three independent experiments, presented as means ± standard deviation (SD). Immunoblot graphs are representative of three independent experiments. Measures of statistical significance were determined by one-way ANOVA for the different amounts of beads versus 4 × 10 6 beads and displayed as: ns; * P

    Journal: BMC Biology

    Article Title: Molecular insights into the surface-specific arrangement of complement C5 convertase enzymes

    doi: 10.1186/s12915-015-0203-8

    Figure Lengend Snippet: High surface density of C3b is critical for C5 convertase activity. a Preparation of beads with different densities of C3b. b Mixing a fixed amount of C3b molecules with increasing numbers of streptavidin beads results in lower C3b densities per bead (top, flow cytometry) while total levels of C3b per sample are equal (below, immunoblot). c C5 convertase activity on streptavidin beads with different C3b densities. d C5 convertase activity plotted against the absolute number of C3b molecules per μm 2 (calculated from the results in c ). b – d Data of three independent experiments, presented as means ± standard deviation (SD). Immunoblot graphs are representative of three independent experiments. Measures of statistical significance were determined by one-way ANOVA for the different amounts of beads versus 4 × 10 6 beads and displayed as: ns; * P

    Article Snippet: Preparation of C3b-coated beads Streptavidin-coated magnetic beads (Dynabeads M-270 Streptavidin, Invitrogen (Carlsbad, CA, USA), 2.8 μm diameter) were washed twice and suspended in VBS-T+ (veronal buffered saline: 2 mM veronal, 145 mM NaCl, pH 7.4 (VBS, pH 7.4) containing 2.5 mM MgCl2 and 0.05 % Tween).

    Techniques: Activity Assay, Flow Cytometry, Cytometry, Standard Deviation

    Attachment of C3b with the thioester toward the surface is critical for C5 convertase activity. a Left, streptavidin beads with site-specifically biotinylated C3b molecules. Right, self-amplified C3b beads were generated by coating streptavidin beads with a low concentration of C3b-biotin after which FB, FD and C3 were added for five repeating incubations to allow natural deposition of C3b and formation of covalently associated C3b multimers (outlined in red). b C5 convertase activity on self-amplified and biotinylated C3b beads. Beads (containing equal levels of C3b) were incubated with FB, FD and C5 and C5a release was determined by calcium mobilization. c Random C3b beads were generated by coupling C3b-biotin onto tosyl-activated beads. d C5 convertase activity on random and biotinylated C3b beads. (b, d) Data of three independent experiments, presented as means ± standard deviation (SD)

    Journal: BMC Biology

    Article Title: Molecular insights into the surface-specific arrangement of complement C5 convertase enzymes

    doi: 10.1186/s12915-015-0203-8

    Figure Lengend Snippet: Attachment of C3b with the thioester toward the surface is critical for C5 convertase activity. a Left, streptavidin beads with site-specifically biotinylated C3b molecules. Right, self-amplified C3b beads were generated by coating streptavidin beads with a low concentration of C3b-biotin after which FB, FD and C3 were added for five repeating incubations to allow natural deposition of C3b and formation of covalently associated C3b multimers (outlined in red). b C5 convertase activity on self-amplified and biotinylated C3b beads. Beads (containing equal levels of C3b) were incubated with FB, FD and C5 and C5a release was determined by calcium mobilization. c Random C3b beads were generated by coupling C3b-biotin onto tosyl-activated beads. d C5 convertase activity on random and biotinylated C3b beads. (b, d) Data of three independent experiments, presented as means ± standard deviation (SD)

    Article Snippet: Preparation of C3b-coated beads Streptavidin-coated magnetic beads (Dynabeads M-270 Streptavidin, Invitrogen (Carlsbad, CA, USA), 2.8 μm diameter) were washed twice and suspended in VBS-T+ (veronal buffered saline: 2 mM veronal, 145 mM NaCl, pH 7.4 (VBS, pH 7.4) containing 2.5 mM MgCl2 and 0.05 % Tween).

    Techniques: Activity Assay, Amplification, Generated, Concentration Assay, Incubation, Standard Deviation