fluorescent snap substrate  (New England Biolabs)


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    Name:
    SNAP Cell TMR STAR
    Description:
    SNAP Cell TMR STAR 30 nmol
    Catalog Number:
    S9105S
    Price:
    335
    Size:
    30 nmol
    Category:
    Fluorochromes
    Score:
    85
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    Structured Review

    New England Biolabs fluorescent snap substrate
    SNAP Cell TMR STAR
    SNAP Cell TMR STAR 30 nmol
    https://www.bioz.com/result/fluorescent snap substrate/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fluorescent snap substrate - by Bioz Stars, 2019-12
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Clathrin promotes centrosome integrity in early mitosis through stabilization of centrosomal ch-TOG
    Article Snippet: For in vivo labeling of SNAP-tagged uLCa fusion proteins, live cells plated on coverslips were incubated (30 min at 37°C in 5% CO2 ) with 3 µM SNAP-TMR-Star (New England Biolabs, Inc.) diluted in normal growth media with serum. .. Covalent cross-linking of SNAP-uLCa was achieved using BG-GLA-BG, a SNAP-tag homodimerizer shown in Fig. S5 , synthesized at New England Biolabs, Inc. following established methods ( ).

    Centrifugation:

    Article Title: Genome-Wide Analysis of Cell Type-Specific Gene Transcription during Spore Formation in Clostridium difficileThe Spore Differentiation Pathway in the Enteric Pathogen Clostridium difficile
    Article Snippet: 1 ml samples were withdrawn from SM cultures at the desired times following inoculation, and the cells collected by centrifugation (4000 g , 10 min, at 4°C). .. For SNAP staining, 1 ml samples were stained for 30 min with 50 nM SNAP-Cell TMR-Star (New England Biolabs) as described .

    Article Title: A Recombination Directionality Factor Controls the Cell Type-Specific Activation of σK and the Fidelity of Spore Development in Clostridium difficile
    Article Snippet: Samples (1 ml) were withdrawn from SM cultures at the desired times following inoculation, and the cells collected by centrifugation (4000 xg , for 10 min, at 4°C). .. For SNAP staining, culture samples of 1 ml were stained for 30 min with 250 nM SNAP-Cell TMR-Star (New England Biolabs) as described before [ ].

    Mass Spectrometry:

    Article Title: The F-actin modifier villin regulates insulin granule dynamics and exocytosis downstream of islet cell autoantigen 512
    Article Snippet: Insulin-SNAP+ SGs were labeled as previously described with the SNAP substrate TMR-Star (New England Biolabs, Ipswich, MA). .. Insulin-SNAP+ SGs were labeled as previously described with the SNAP substrate TMR-Star (New England Biolabs, Ipswich, MA).

    Stable Transfection:

    Article Title: Comparison of a mouse and a novel human scFv-SNAP-auristatin F drug conjugate with potent activity against EGFR-overexpressing human solid tumor cells
    Article Snippet: The effects on microtubule dynamics were visualized as previously described, using A549 cells stably expressing a recombinant SNAP-TubB3. .. The recombinant tubulin structure was stained directly by adding 50 µL/well of a solution in growth medium containing 30 nM SNAP-Cell TMR-Star (NEB) for labeling the tubulin and 1 µg/mL 4′,6-diamidino-2-phenylindole (Thermo Fisher Scientific) for nuclei counterstaining.

    Article Title: POLE3-POLE4 Is a Histone H3-H4 Chaperone that Maintains Chromatin Integrity during DNA Replication
    Article Snippet: HeLa cells stably expressing H3.1-SNAP-3xHA were previously characterized ( ). .. Finally, we performed a pulse step by incubating cells for 20 min in complete medium containing 2 μM of SNAP-Cell TMR-Star (New England Biolabs) and 10 μM of EdU (5-ethynyl-20-deoxyuridine).

    Article Title: Lipids Regulate Lck Protein Activity through Their Interactions with the Lck Src Homology 2 Domain
    Article Snippet: Simultaneous single molecule imaging of two proteins was performed in Jurkat cells using a total internal reflection fluorescence microscope as described previously ( ). .. JCaM1.6 Jurkat cells stably expressing Lck-GFP WT (or mutants) were transiently transfected with SNAP®-tagged TCR-ζ chain, which was subsequently labeled with SNAP-Cell® TMR-Star® (New England Biolabs). .. Labeled cells were washed, attached to a poly- l -lysine-coated glass plate for imaging, and stimulated with 10 μg/ml OKT3, and Lck and ζ molecules were simultaneously tracked.

    Synthesized:

    Article Title: A novel role for the histone acetyltransferase Hat1 in the CENP-A/CID assembly pathway in Drosophila melanogaster
    Article Snippet: Paragraph title: Quench-chase-pulse labeling of newly synthesized CENP-A ... After 48 h (Figure , Supplementary Figure S6) or 24 h (Supplementary Figure S6) chase, cells were pulse-labeled with 4.5 μM SNAP-Cell TMR Star (NEB) for 30 min. Nonreacted TMR Star was washed out, cells were fixed in 3.7% paraformaldehyde/0.3% Triton-X100 and nuclei were counterstained with DAPI.

    Article Title: Clathrin promotes centrosome integrity in early mitosis through stabilization of centrosomal ch-TOG
    Article Snippet: For in vivo labeling of SNAP-tagged uLCa fusion proteins, live cells plated on coverslips were incubated (30 min at 37°C in 5% CO2 ) with 3 µM SNAP-TMR-Star (New England Biolabs, Inc.) diluted in normal growth media with serum. .. Cells were then washed three times with normal growth media with serum, incubated (37°C in 5% CO2 ) an additional 30 min, and then imaged using confocal laser-scanning microscopy, as detailed under Time-lapse confocal microscopy imaging.

    Blocking Assay:

    Article Title: POLE3-POLE4 Is a Histone H3-H4 Chaperone that Maintains Chromatin Integrity during DNA Replication
    Article Snippet: For quench-chase-pulse experiments, cells were incubated in complete medium containing 10 μM of SNAP-Cell Block (New England Biolabs) to quench SNAP tag activity, washed twice with PBS, and incubated for 30 min in complete medium to allow SNAP-Cell Block to diffuse out. .. Finally, we performed a pulse step by incubating cells for 20 min in complete medium containing 2 μM of SNAP-Cell TMR-Star (New England Biolabs) and 10 μM of EdU (5-ethynyl-20-deoxyuridine).

    Article Title: A novel role for the histone acetyltransferase Hat1 in the CENP-A/CID assembly pathway in Drosophila melanogaster
    Article Snippet: Quenching of SNAP tag activity of existing SNAP-CENP-A was performed by a 30 min incubation of cells with 5 μM SNAP-Cell Block (NEB). .. After 48 h (Figure , Supplementary Figure S6) or 24 h (Supplementary Figure S6) chase, cells were pulse-labeled with 4.5 μM SNAP-Cell TMR Star (NEB) for 30 min. Nonreacted TMR Star was washed out, cells were fixed in 3.7% paraformaldehyde/0.3% Triton-X100 and nuclei were counterstained with DAPI.

    Article Title: BLOC-1 is required for selective membrane protein trafficking from endosomes to primary cilia
    Article Snippet: Cells were washed with 1× PBS and blocked in 1% BSA in 1× TBST (150 mM NaCl, 1% Tween 20, and 50 mM Tris, pH 7.5) for 1 h. Primary antibodies were diluted in blocking solution and incubated for 2 h at room temperature. .. Cells were washed four times with blocking solution over 20 min. Next, they were incubated with 1:2,000 dilutions of either Alexa Fluor 488–, 568–, 594–, 647–, and 680–conjugated IgG anti-mouse or IgG anti-rabbit (Thermo Fisher Scientific), SNAP TMR STAR (New England Biolabs, Inc.) for 1 h. Cells were then washed again with blocking solution, followed by three washes with 1× PBS. .. Coverslips were mounted onto slides with prolong medium (Molecular Probes).

    Incubation:

    Article Title: A SNAP-Tagged Derivative of HIV-1--A Versatile Tool to Study Virus-Cell Interactions
    Article Snippet: Signals were detected by quantitative immunoblot using a LiCor Odyssey system as well as reagents and protocols provided by the manufacturer and quantitated using Odyssey 2.1 software. .. Hela cells seeded on coverslips 24 h prior to the experiment were transfected with FuGene6 (Roche) and incubated at 37°C for 24 h. For SNAP-labelling cells were incubated with 1 µM SNAP-cell TMR-Star (New England Biolabs) in DMEM for 15 min at 37°C, washed and incubated in fresh DMEM for 45 min at 37°C to allow diffusion of excess substrate. .. Cells were then fixed with 3% paraformaldehyde (PFA), counterstained with Hoechst 33258 and embedded in Mowiol.

    Article Title: Lissencephaly-1 is a context-dependent regulator of the human dynein complex
    Article Snippet: Protein concentrations were measured using the Coomassie Protein Assay Kit (ThermoFisher Scientific) and an Eppendorf BioPhotometer Plus (Eppendorf (Hamburg, Germany)). .. IgG Sepharose 6 Fast Flow beads bound to SNAPf ::dynein complexes or LIS1::SNAPf were incubated at 4°C for 40 min with ~5 μM SNAP-Surface Alexa Fluor 647 or SNAP-Cell TMR-Star (NEB) for SNAPf ::dynein, or ~40 μM SNAP-Cell TMR-Star for LIS1::SNAPf . .. Prior to TEV cleavage, excess dye was removed with three washes (25 ml each for dynein; 2 ml each for LIS1) in TEV Buffer, and beads resuspended in 300 μl TEV buffer.

    Article Title: Comparison of a mouse and a novel human scFv-SNAP-auristatin F drug conjugate with potent activity against EGFR-overexpressing human solid tumor cells
    Article Snippet: The recombinant tubulin structure was stained directly by adding 50 µL/well of a solution in growth medium containing 30 nM SNAP-Cell TMR-Star (NEB) for labeling the tubulin and 1 µg/mL 4′,6-diamidino-2-phenylindole (Thermo Fisher Scientific) for nuclei counterstaining. .. The cells were incubated overnight at 37°C, 5% CO2 and 100% humidity, before incubation with 50, 150, or 250 nM of the EGFR-specific ADCs.

    Article Title: Molecular coordination of Staphylococcus aureus cell division
    Article Snippet: The volume was then calculated based on a prolate spheroid shape with volume V = 4 3 π a b 2 , where a and b are the dimensions of the long and short axis respectively. .. SNAP-Cell TMR-Star (New England Biolabs) was added to a 1 ml aliquot of mid-exponential phase (OD600 ~1) grown culture at a concentration of 500 nM and incubated at 37°C for 1 hr. .. Cells were washed three times by resuspension and centrifugation in PBS, resuspended in PBS supplemented with 200 μg ml−1 lysostaphin and 20 U ml−1 DNase I and lysed at 37°C for 30 min.

    Article Title: POLE3-POLE4 Is a Histone H3-H4 Chaperone that Maintains Chromatin Integrity during DNA Replication
    Article Snippet: For quench-chase-pulse experiments, cells were incubated in complete medium containing 10 μM of SNAP-Cell Block (New England Biolabs) to quench SNAP tag activity, washed twice with PBS, and incubated for 30 min in complete medium to allow SNAP-Cell Block to diffuse out. .. Finally, we performed a pulse step by incubating cells for 20 min in complete medium containing 2 μM of SNAP-Cell TMR-Star (New England Biolabs) and 10 μM of EdU (5-ethynyl-20-deoxyuridine).

    Article Title: A novel role for the histone acetyltransferase Hat1 in the CENP-A/CID assembly pathway in Drosophila melanogaster
    Article Snippet: Quenching of SNAP tag activity of existing SNAP-CENP-A was performed by a 30 min incubation of cells with 5 μM SNAP-Cell Block (NEB). .. After 48 h (Figure , Supplementary Figure S6) or 24 h (Supplementary Figure S6) chase, cells were pulse-labeled with 4.5 μM SNAP-Cell TMR Star (NEB) for 30 min. Nonreacted TMR Star was washed out, cells were fixed in 3.7% paraformaldehyde/0.3% Triton-X100 and nuclei were counterstained with DAPI.

    Article Title: Molecular coordination of Staphylococcus aureus cell division
    Article Snippet: Cells were then washed by centrifugation and resuspension in PBS. .. S. aureus grown to mid-exponential phase (OD600 ~0.5) were incubated with SNAP-Cell TMR-Star (New England Biolabs) at 500 nM for widefield microscopy or 3 μM for SIM at 37°C for 15 min. .. Cells were washed by centrifugation and resuspension in PBS.

    Article Title: Live-cell protein labelling with nanometre precision by cell squeezing
    Article Snippet: As a control for endosomal uptake, cells were incubated with 100 nM of tris NTAf at RT and 4 °C without microfluidic cell manipulation ( ). .. In case of combined tris NTAf and SNAPf -tag labelling, cells were first squeezed in the presence of 100 nM tris NTAAlexa647 and 3 μM SNAP-Cell TMR-Star (New England Biolabs) was added 5 min after squeezing.

    Article Title: Clathrin promotes centrosome integrity in early mitosis through stabilization of centrosomal ch-TOG
    Article Snippet: Precleared and IP samples were resolved by SDS-PAGE, transferred to nitrocellulose membranes, and analyzed by immunoblotting. .. For in vivo labeling of SNAP-tagged uLCa fusion proteins, live cells plated on coverslips were incubated (30 min at 37°C in 5% CO2 ) with 3 µM SNAP-TMR-Star (New England Biolabs, Inc.) diluted in normal growth media with serum. .. Cells were then washed three times with normal growth media with serum, incubated (37°C in 5% CO2 ) an additional 30 min, and then imaged using confocal laser-scanning microscopy, as detailed under Time-lapse confocal microscopy imaging.

    Article Title: BLOC-1 is required for selective membrane protein trafficking from endosomes to primary cilia
    Article Snippet: Cells were washed with 1× PBS and blocked in 1% BSA in 1× TBST (150 mM NaCl, 1% Tween 20, and 50 mM Tris, pH 7.5) for 1 h. Primary antibodies were diluted in blocking solution and incubated for 2 h at room temperature. .. Cells were washed four times with blocking solution over 20 min. Next, they were incubated with 1:2,000 dilutions of either Alexa Fluor 488–, 568–, 594–, 647–, and 680–conjugated IgG anti-mouse or IgG anti-rabbit (Thermo Fisher Scientific), SNAP TMR STAR (New England Biolabs, Inc.) for 1 h. Cells were then washed again with blocking solution, followed by three washes with 1× PBS. .. Coverslips were mounted onto slides with prolong medium (Molecular Probes).

    Article Title: The F-actin modifier villin regulates insulin granule dynamics and exocytosis downstream of islet cell autoantigen 512
    Article Snippet: 2.9 Prior to imaging, cells grown in an open chamber were incubated in resting media and transferred onto a thermostat-controlled (37 °C) stage. .. Insulin-SNAP+ SGs were labeled as previously described with the SNAP substrate TMR-Star (New England Biolabs, Ipswich, MA).

    Article Title:
    Article Snippet: After 10 min, unattached axonemes were washed away with HMEEK buffer. .. The cover glasses were incubated with a droplet of 3 μ m SNAP tag BG-substrate (which forms a covalent bond with the SNAP tag) SNAP-Cell® TMR Star (New England Biolabs, Ipswich, MA) in HMEEK buffer with 1% BSA for 2 h at room temperature. .. Excess SNAP-Cell® TMR Star was removed by three washing steps with HMEEK buffer.

    Diffusion-based Assay:

    Article Title: A SNAP-Tagged Derivative of HIV-1--A Versatile Tool to Study Virus-Cell Interactions
    Article Snippet: Signals were detected by quantitative immunoblot using a LiCor Odyssey system as well as reagents and protocols provided by the manufacturer and quantitated using Odyssey 2.1 software. .. Hela cells seeded on coverslips 24 h prior to the experiment were transfected with FuGene6 (Roche) and incubated at 37°C for 24 h. For SNAP-labelling cells were incubated with 1 µM SNAP-cell TMR-Star (New England Biolabs) in DMEM for 15 min at 37°C, washed and incubated in fresh DMEM for 45 min at 37°C to allow diffusion of excess substrate. .. Cells were then fixed with 3% paraformaldehyde (PFA), counterstained with Hoechst 33258 and embedded in Mowiol.

    Activity Assay:

    Article Title: POLE3-POLE4 Is a Histone H3-H4 Chaperone that Maintains Chromatin Integrity during DNA Replication
    Article Snippet: For quench-chase-pulse experiments, cells were incubated in complete medium containing 10 μM of SNAP-Cell Block (New England Biolabs) to quench SNAP tag activity, washed twice with PBS, and incubated for 30 min in complete medium to allow SNAP-Cell Block to diffuse out. .. Finally, we performed a pulse step by incubating cells for 20 min in complete medium containing 2 μM of SNAP-Cell TMR-Star (New England Biolabs) and 10 μM of EdU (5-ethynyl-20-deoxyuridine).

    Article Title: A novel role for the histone acetyltransferase Hat1 in the CENP-A/CID assembly pathway in Drosophila melanogaster
    Article Snippet: Quenching of SNAP tag activity of existing SNAP-CENP-A was performed by a 30 min incubation of cells with 5 μM SNAP-Cell Block (NEB). .. After 48 h (Figure , Supplementary Figure S6) or 24 h (Supplementary Figure S6) chase, cells were pulse-labeled with 4.5 μM SNAP-Cell TMR Star (NEB) for 30 min. Nonreacted TMR Star was washed out, cells were fixed in 3.7% paraformaldehyde/0.3% Triton-X100 and nuclei were counterstained with DAPI.

    Infection:

    Article Title: A SNAP-Tagged Derivative of HIV-1--A Versatile Tool to Study Virus-Cell Interactions
    Article Snippet: Hela cells seeded on coverslips 24 h prior to the experiment were transfected with FuGene6 (Roche) and incubated at 37°C for 24 h. For SNAP-labelling cells were incubated with 1 µM SNAP-cell TMR-Star (New England Biolabs) in DMEM for 15 min at 37°C, washed and incubated in fresh DMEM for 45 min at 37°C to allow diffusion of excess substrate. .. Hela cells seeded on coverslips 24 h prior to the experiment were transfected with FuGene6 (Roche) and incubated at 37°C for 24 h. For SNAP-labelling cells were incubated with 1 µM SNAP-cell TMR-Star (New England Biolabs) in DMEM for 15 min at 37°C, washed and incubated in fresh DMEM for 45 min at 37°C to allow diffusion of excess substrate.

    Expressing:

    Article Title: Comparison of a mouse and a novel human scFv-SNAP-auristatin F drug conjugate with potent activity against EGFR-overexpressing human solid tumor cells
    Article Snippet: The effects on microtubule dynamics were visualized as previously described, using A549 cells stably expressing a recombinant SNAP-TubB3. .. The recombinant tubulin structure was stained directly by adding 50 µL/well of a solution in growth medium containing 30 nM SNAP-Cell TMR-Star (NEB) for labeling the tubulin and 1 µg/mL 4′,6-diamidino-2-phenylindole (Thermo Fisher Scientific) for nuclei counterstaining.

    Article Title: POLE3-POLE4 Is a Histone H3-H4 Chaperone that Maintains Chromatin Integrity during DNA Replication
    Article Snippet: HeLa cells stably expressing H3.1-SNAP-3xHA were previously characterized ( ). .. Finally, we performed a pulse step by incubating cells for 20 min in complete medium containing 2 μM of SNAP-Cell TMR-Star (New England Biolabs) and 10 μM of EdU (5-ethynyl-20-deoxyuridine).

    Article Title: Lipids Regulate Lck Protein Activity through Their Interactions with the Lck Src Homology 2 Domain
    Article Snippet: Simultaneous single molecule imaging of two proteins was performed in Jurkat cells using a total internal reflection fluorescence microscope as described previously ( ). .. JCaM1.6 Jurkat cells stably expressing Lck-GFP WT (or mutants) were transiently transfected with SNAP®-tagged TCR-ζ chain, which was subsequently labeled with SNAP-Cell® TMR-Star® (New England Biolabs). .. Labeled cells were washed, attached to a poly- l -lysine-coated glass plate for imaging, and stimulated with 10 μg/ml OKT3, and Lck and ζ molecules were simultaneously tracked.

    Modification:

    Article Title: A SNAP-Tagged Derivative of HIV-1--A Versatile Tool to Study Virus-Cell Interactions
    Article Snippet: Hela cells seeded on coverslips 24 h prior to the experiment were transfected with FuGene6 (Roche) and incubated at 37°C for 24 h. For SNAP-labelling cells were incubated with 1 µM SNAP-cell TMR-Star (New England Biolabs) in DMEM for 15 min at 37°C, washed and incubated in fresh DMEM for 45 min at 37°C to allow diffusion of excess substrate. .. Hela cells seeded on coverslips 24 h prior to the experiment were transfected with FuGene6 (Roche) and incubated at 37°C for 24 h. For SNAP-labelling cells were incubated with 1 µM SNAP-cell TMR-Star (New England Biolabs) in DMEM for 15 min at 37°C, washed and incubated in fresh DMEM for 45 min at 37°C to allow diffusion of excess substrate.

    MANN-WHITNEY:

    Article Title: A novel role for the histone acetyltransferase Hat1 in the CENP-A/CID assembly pathway in Drosophila melanogaster
    Article Snippet: After 48 h (Figure , Supplementary Figure S6) or 24 h (Supplementary Figure S6) chase, cells were pulse-labeled with 4.5 μM SNAP-Cell TMR Star (NEB) for 30 min. Nonreacted TMR Star was washed out, cells were fixed in 3.7% paraformaldehyde/0.3% Triton-X100 and nuclei were counterstained with DAPI. .. After 48 h (Figure , Supplementary Figure S6) or 24 h (Supplementary Figure S6) chase, cells were pulse-labeled with 4.5 μM SNAP-Cell TMR Star (NEB) for 30 min. Nonreacted TMR Star was washed out, cells were fixed in 3.7% paraformaldehyde/0.3% Triton-X100 and nuclei were counterstained with DAPI.

    Inhibition:

    Article Title: Clathrin promotes centrosome integrity in early mitosis through stabilization of centrosomal ch-TOG
    Article Snippet: Paragraph title: SNAP-tag labeling, fusion protein cross-linking, and Aurora A kinase inhibition assays ... For in vivo labeling of SNAP-tagged uLCa fusion proteins, live cells plated on coverslips were incubated (30 min at 37°C in 5% CO2 ) with 3 µM SNAP-TMR-Star (New England Biolabs, Inc.) diluted in normal growth media with serum.

    Transfection:

    Article Title: A SNAP-Tagged Derivative of HIV-1--A Versatile Tool to Study Virus-Cell Interactions
    Article Snippet: Signals were detected by quantitative immunoblot using a LiCor Odyssey system as well as reagents and protocols provided by the manufacturer and quantitated using Odyssey 2.1 software. .. Hela cells seeded on coverslips 24 h prior to the experiment were transfected with FuGene6 (Roche) and incubated at 37°C for 24 h. For SNAP-labelling cells were incubated with 1 µM SNAP-cell TMR-Star (New England Biolabs) in DMEM for 15 min at 37°C, washed and incubated in fresh DMEM for 45 min at 37°C to allow diffusion of excess substrate. .. Cells were then fixed with 3% paraformaldehyde (PFA), counterstained with Hoechst 33258 and embedded in Mowiol.

    Article Title: Lipids Regulate Lck Protein Activity through Their Interactions with the Lck Src Homology 2 Domain
    Article Snippet: Simultaneous single molecule imaging of two proteins was performed in Jurkat cells using a total internal reflection fluorescence microscope as described previously ( ). .. JCaM1.6 Jurkat cells stably expressing Lck-GFP WT (or mutants) were transiently transfected with SNAP®-tagged TCR-ζ chain, which was subsequently labeled with SNAP-Cell® TMR-Star® (New England Biolabs). .. Labeled cells were washed, attached to a poly- l -lysine-coated glass plate for imaging, and stimulated with 10 μg/ml OKT3, and Lck and ζ molecules were simultaneously tracked.

    Pulse Chase:

    Article Title: POLE3-POLE4 Is a Histone H3-H4 Chaperone that Maintains Chromatin Integrity during DNA Replication
    Article Snippet: Finally, we performed a pulse step by incubating cells for 20 min in complete medium containing 2 μM of SNAP-Cell TMR-Star (New England Biolabs) and 10 μM of EdU (5-ethynyl-20-deoxyuridine). .. Finally, we performed a pulse step by incubating cells for 20 min in complete medium containing 2 μM of SNAP-Cell TMR-Star (New England Biolabs) and 10 μM of EdU (5-ethynyl-20-deoxyuridine).

    Cell Culture:

    Article Title: Comparison of a mouse and a novel human scFv-SNAP-auristatin F drug conjugate with potent activity against EGFR-overexpressing human solid tumor cells
    Article Snippet: Briefly, 7×103 cells were plated in a flat bottom black 96-well half-area microplate (µclear; Greiner Bio-One) in a total volume of 50 µL cell culture medium without G418. .. The recombinant tubulin structure was stained directly by adding 50 µL/well of a solution in growth medium containing 30 nM SNAP-Cell TMR-Star (NEB) for labeling the tubulin and 1 µg/mL 4′,6-diamidino-2-phenylindole (Thermo Fisher Scientific) for nuclei counterstaining.

    Article Title: Live-cell protein labelling with nanometre precision by cell squeezing
    Article Snippet: Squeezed cells were washed with DMEM containing 10% FCS and 10 mM histidine (Sigma-Aldrich), to remove unspecifically bound tris NTAf from the cell surface, seeded into eight wells on cover glass II slides (Sarstedt) in DMEM containing 10% FCS and cultured at 37 °C and 5% CO2 . .. In case of combined tris NTAf and SNAPf -tag labelling, cells were first squeezed in the presence of 100 nM tris NTAAlexa647 and 3 μM SNAP-Cell TMR-Star (New England Biolabs) was added 5 min after squeezing.

    Light Microscopy:

    Article Title:
    Article Snippet: Paragraph title: Fluorescence Light Microscopy ... The cover glasses were incubated with a droplet of 3 μ m SNAP tag BG-substrate (which forms a covalent bond with the SNAP tag) SNAP-Cell® TMR Star (New England Biolabs, Ipswich, MA) in HMEEK buffer with 1% BSA for 2 h at room temperature.

    other:

    Article Title: RNA-directed activation of cytoplasmic dynein-1 in reconstituted transport RNPs
    Article Snippet: The ratio of SNAP-Surface Alexa Fluor 647 to SNAP-Cell TMR-Star that yielded approximately half of labelled polypeptides having one fluorophore and half the other fluorophore was determined empirically for different batches of the dyes.

    Imaging:

    Article Title: A Recombination Directionality Factor Controls the Cell Type-Specific Activation of σK and the Fidelity of Spore Development in Clostridium difficile
    Article Snippet: For SNAP staining, culture samples of 1 ml were stained for 30 min with 250 nM SNAP-Cell TMR-Star (New England Biolabs) as described before [ ]. .. For SNAP staining, culture samples of 1 ml were stained for 30 min with 250 nM SNAP-Cell TMR-Star (New England Biolabs) as described before [ ].

    Article Title: Lipids Regulate Lck Protein Activity through Their Interactions with the Lck Src Homology 2 Domain
    Article Snippet: Simultaneous single molecule imaging of two proteins was performed in Jurkat cells using a total internal reflection fluorescence microscope as described previously ( ). .. JCaM1.6 Jurkat cells stably expressing Lck-GFP WT (or mutants) were transiently transfected with SNAP®-tagged TCR-ζ chain, which was subsequently labeled with SNAP-Cell® TMR-Star® (New England Biolabs).

    Article Title: Live-cell protein labelling with nanometre precision by cell squeezing
    Article Snippet: In case of combined tris NTAf and SNAPf -tag labelling, cells were first squeezed in the presence of 100 nM tris NTAAlexa647 and 3 μM SNAP-Cell TMR-Star (New England Biolabs) was added 5 min after squeezing. .. In case of combined tris NTAf and SNAPf -tag labelling, cells were first squeezed in the presence of 100 nM tris NTAAlexa647 and 3 μM SNAP-Cell TMR-Star (New England Biolabs) was added 5 min after squeezing.

    Article Title: The F-actin modifier villin regulates insulin granule dynamics and exocytosis downstream of islet cell autoantigen 512
    Article Snippet: 2.9 Prior to imaging, cells grown in an open chamber were incubated in resting media and transferred onto a thermostat-controlled (37 °C) stage. .. Insulin-SNAP+ SGs were labeled as previously described with the SNAP substrate TMR-Star (New England Biolabs, Ipswich, MA).

    Article Title:
    Article Snippet: The cover glasses were incubated with a droplet of 3 μ m SNAP tag BG-substrate (which forms a covalent bond with the SNAP tag) SNAP-Cell® TMR Star (New England Biolabs, Ipswich, MA) in HMEEK buffer with 1% BSA for 2 h at room temperature. .. In the first washing step, the cover glasses were incubated with HMEEK buffer for 30 min to improve the washing.

    Recombinant:

    Article Title: Comparison of a mouse and a novel human scFv-SNAP-auristatin F drug conjugate with potent activity against EGFR-overexpressing human solid tumor cells
    Article Snippet: Briefly, 7×103 cells were plated in a flat bottom black 96-well half-area microplate (µclear; Greiner Bio-One) in a total volume of 50 µL cell culture medium without G418. .. The recombinant tubulin structure was stained directly by adding 50 µL/well of a solution in growth medium containing 30 nM SNAP-Cell TMR-Star (NEB) for labeling the tubulin and 1 µg/mL 4′,6-diamidino-2-phenylindole (Thermo Fisher Scientific) for nuclei counterstaining. .. The cells were incubated overnight at 37°C, 5% CO2 and 100% humidity, before incubation with 50, 150, or 250 nM of the EGFR-specific ADCs.

    Article Title: Tyrosination of α‐tubulin controls the initiation of processive dynein–dynactin motility
    Article Snippet: The DDB complex was prepared by adding recombinant strepII‐SNAPf‐tagged BiCD2 (N‐terminal construct encompassing amino acids 25–400) to high‐speed porcine brain lysates as previously described (McKenney et al , ). .. The DDB complexes were fluorescently labeled with excess SNAP‐Cell TMR‐Star dye (NEB) during purification as described (McKenney et al , ), and aliquots of eluted DDB were flash‐frozen in LN2 and stored at −80°C.

    Immunofluorescence:

    Article Title: BLOC-1 is required for selective membrane protein trafficking from endosomes to primary cilia
    Article Snippet: Paragraph title: Immunofluorescence ... Cells were washed four times with blocking solution over 20 min. Next, they were incubated with 1:2,000 dilutions of either Alexa Fluor 488–, 568–, 594–, 647–, and 680–conjugated IgG anti-mouse or IgG anti-rabbit (Thermo Fisher Scientific), SNAP TMR STAR (New England Biolabs, Inc.) for 1 h. Cells were then washed again with blocking solution, followed by three washes with 1× PBS.

    In Vivo:

    Article Title: POLE3-POLE4 Is a Histone H3-H4 Chaperone that Maintains Chromatin Integrity during DNA Replication
    Article Snippet: Paragraph title: H3.1-SNAP assay in vivo ... Finally, we performed a pulse step by incubating cells for 20 min in complete medium containing 2 μM of SNAP-Cell TMR-Star (New England Biolabs) and 10 μM of EdU (5-ethynyl-20-deoxyuridine).

    Article Title: Clathrin promotes centrosome integrity in early mitosis through stabilization of centrosomal ch-TOG
    Article Snippet: Precleared and IP samples were resolved by SDS-PAGE, transferred to nitrocellulose membranes, and analyzed by immunoblotting. .. For in vivo labeling of SNAP-tagged uLCa fusion proteins, live cells plated on coverslips were incubated (30 min at 37°C in 5% CO2 ) with 3 µM SNAP-TMR-Star (New England Biolabs, Inc.) diluted in normal growth media with serum. .. Cells were then washed three times with normal growth media with serum, incubated (37°C in 5% CO2 ) an additional 30 min, and then imaged using confocal laser-scanning microscopy, as detailed under Time-lapse confocal microscopy imaging.

    Fluorescence:

    Article Title: Lipids Regulate Lck Protein Activity through Their Interactions with the Lck Src Homology 2 Domain
    Article Snippet: Paragraph title: Single Molecule Tracking by Total Internal Reflection Fluorescence Microscopy ... JCaM1.6 Jurkat cells stably expressing Lck-GFP WT (or mutants) were transiently transfected with SNAP®-tagged TCR-ζ chain, which was subsequently labeled with SNAP-Cell® TMR-Star® (New England Biolabs).

    Article Title: The F-actin modifier villin regulates insulin granule dynamics and exocytosis downstream of islet cell autoantigen 512
    Article Snippet: Paragraph title: Total internal reflection fluorescence microscopy (TIRFM) ... Insulin-SNAP+ SGs were labeled as previously described with the SNAP substrate TMR-Star (New England Biolabs, Ipswich, MA).

    Article Title:
    Article Snippet: Paragraph title: Fluorescence Light Microscopy ... The cover glasses were incubated with a droplet of 3 μ m SNAP tag BG-substrate (which forms a covalent bond with the SNAP tag) SNAP-Cell® TMR Star (New England Biolabs, Ipswich, MA) in HMEEK buffer with 1% BSA for 2 h at room temperature.

    Isolation:

    Article Title:
    Article Snippet: The cover glasses were incubated with a droplet of 3 μ m SNAP tag BG-substrate (which forms a covalent bond with the SNAP tag) SNAP-Cell® TMR Star (New England Biolabs, Ipswich, MA) in HMEEK buffer with 1% BSA for 2 h at room temperature. .. The cover glasses were incubated with a droplet of 3 μ m SNAP tag BG-substrate (which forms a covalent bond with the SNAP tag) SNAP-Cell® TMR Star (New England Biolabs, Ipswich, MA) in HMEEK buffer with 1% BSA for 2 h at room temperature.

    Flow Cytometry:

    Article Title: Lissencephaly-1 is a context-dependent regulator of the human dynein complex
    Article Snippet: Protein concentrations were measured using the Coomassie Protein Assay Kit (ThermoFisher Scientific) and an Eppendorf BioPhotometer Plus (Eppendorf (Hamburg, Germany)). .. IgG Sepharose 6 Fast Flow beads bound to SNAPf ::dynein complexes or LIS1::SNAPf were incubated at 4°C for 40 min with ~5 μM SNAP-Surface Alexa Fluor 647 or SNAP-Cell TMR-Star (NEB) for SNAPf ::dynein, or ~40 μM SNAP-Cell TMR-Star for LIS1::SNAPf . .. Prior to TEV cleavage, excess dye was removed with three washes (25 ml each for dynein; 2 ml each for LIS1) in TEV Buffer, and beads resuspended in 300 μl TEV buffer.

    Microscopy:

    Article Title: Comparison of a mouse and a novel human scFv-SNAP-auristatin F drug conjugate with potent activity against EGFR-overexpressing human solid tumor cells
    Article Snippet: The recombinant tubulin structure was stained directly by adding 50 µL/well of a solution in growth medium containing 30 nM SNAP-Cell TMR-Star (NEB) for labeling the tubulin and 1 µg/mL 4′,6-diamidino-2-phenylindole (Thermo Fisher Scientific) for nuclei counterstaining. .. The cells were incubated overnight at 37°C, 5% CO2 and 100% humidity, before incubation with 50, 150, or 250 nM of the EGFR-specific ADCs.

    Article Title: Genome-Wide Analysis of Cell Type-Specific Gene Transcription during Spore Formation in Clostridium difficileThe Spore Differentiation Pathway in the Enteric Pathogen Clostridium difficile
    Article Snippet: Paragraph title: Microscopy and image analysis ... For SNAP staining, 1 ml samples were stained for 30 min with 50 nM SNAP-Cell TMR-Star (New England Biolabs) as described .

    Article Title: A Recombination Directionality Factor Controls the Cell Type-Specific Activation of σK and the Fidelity of Spore Development in Clostridium difficile
    Article Snippet: Paragraph title: Microscopy and image analysis ... For SNAP staining, culture samples of 1 ml were stained for 30 min with 250 nM SNAP-Cell TMR-Star (New England Biolabs) as described before [ ].

    Article Title: Molecular coordination of Staphylococcus aureus cell division
    Article Snippet: Cells were then washed by centrifugation and resuspension in PBS. .. S. aureus grown to mid-exponential phase (OD600 ~0.5) were incubated with SNAP-Cell TMR-Star (New England Biolabs) at 500 nM for widefield microscopy or 3 μM for SIM at 37°C for 15 min. .. Cells were washed by centrifugation and resuspension in PBS.

    Article Title: Lipids Regulate Lck Protein Activity through Their Interactions with the Lck Src Homology 2 Domain
    Article Snippet: Paragraph title: Single Molecule Tracking by Total Internal Reflection Fluorescence Microscopy ... JCaM1.6 Jurkat cells stably expressing Lck-GFP WT (or mutants) were transiently transfected with SNAP®-tagged TCR-ζ chain, which was subsequently labeled with SNAP-Cell® TMR-Star® (New England Biolabs).

    Article Title: BLOC-1 is required for selective membrane protein trafficking from endosomes to primary cilia
    Article Snippet: Cells were washed four times with blocking solution over 20 min. Next, they were incubated with 1:2,000 dilutions of either Alexa Fluor 488–, 568–, 594–, 647–, and 680–conjugated IgG anti-mouse or IgG anti-rabbit (Thermo Fisher Scientific), SNAP TMR STAR (New England Biolabs, Inc.) for 1 h. Cells were then washed again with blocking solution, followed by three washes with 1× PBS. .. Cells were washed four times with blocking solution over 20 min. Next, they were incubated with 1:2,000 dilutions of either Alexa Fluor 488–, 568–, 594–, 647–, and 680–conjugated IgG anti-mouse or IgG anti-rabbit (Thermo Fisher Scientific), SNAP TMR STAR (New England Biolabs, Inc.) for 1 h. Cells were then washed again with blocking solution, followed by three washes with 1× PBS.

    Article Title: The F-actin modifier villin regulates insulin granule dynamics and exocytosis downstream of islet cell autoantigen 512
    Article Snippet: Paragraph title: Total internal reflection fluorescence microscopy (TIRFM) ... Insulin-SNAP+ SGs were labeled as previously described with the SNAP substrate TMR-Star (New England Biolabs, Ipswich, MA).

    Article Title:
    Article Snippet: The cover glasses were incubated with a droplet of 3 μ m SNAP tag BG-substrate (which forms a covalent bond with the SNAP tag) SNAP-Cell® TMR Star (New England Biolabs, Ipswich, MA) in HMEEK buffer with 1% BSA for 2 h at room temperature. .. In the first washing step, the cover glasses were incubated with HMEEK buffer for 30 min to improve the washing.

    Purification:

    Article Title: Tyrosination of α‐tubulin controls the initiation of processive dynein–dynactin motility
    Article Snippet: The DDB complex was prepared by adding recombinant strepII‐SNAPf‐tagged BiCD2 (N‐terminal construct encompassing amino acids 25–400) to high‐speed porcine brain lysates as previously described (McKenney et al , ). .. The DDB complexes were fluorescently labeled with excess SNAP‐Cell TMR‐Star dye (NEB) during purification as described (McKenney et al , ), and aliquots of eluted DDB were flash‐frozen in LN2 and stored at −80°C. .. We note that freezing the complex leads to an apparently larger percentage of diffusive complexes in our assays (~15% for unfrozen versus ~30% for frozen).

    Labeling:

    Article Title: Comparison of a mouse and a novel human scFv-SNAP-auristatin F drug conjugate with potent activity against EGFR-overexpressing human solid tumor cells
    Article Snippet: Briefly, 7×103 cells were plated in a flat bottom black 96-well half-area microplate (µclear; Greiner Bio-One) in a total volume of 50 µL cell culture medium without G418. .. The recombinant tubulin structure was stained directly by adding 50 µL/well of a solution in growth medium containing 30 nM SNAP-Cell TMR-Star (NEB) for labeling the tubulin and 1 µg/mL 4′,6-diamidino-2-phenylindole (Thermo Fisher Scientific) for nuclei counterstaining. .. The cells were incubated overnight at 37°C, 5% CO2 and 100% humidity, before incubation with 50, 150, or 250 nM of the EGFR-specific ADCs.

    Article Title: In Search of Novel Drug Target Sites on Estrogen Receptors Using RNA Aptamers
    Article Snippet: The expression of the recombinant SNAP-ERα was confirmed by fluorescent microscopy. .. Twenty-four hours post-transfection, live HeLa cells were labeled with SNAP-Cell TMR-Star and Hoechst 33342. .. As shown in , the SNAP fusion protein was located in both the cytoplasm and nucleus of transfected cells, but mainly in the nucleus.

    Article Title: A novel role for the histone acetyltransferase Hat1 in the CENP-A/CID assembly pathway in Drosophila melanogaster
    Article Snippet: Paragraph title: Quench-chase-pulse labeling of newly synthesized CENP-A ... After 48 h (Figure , Supplementary Figure S6) or 24 h (Supplementary Figure S6) chase, cells were pulse-labeled with 4.5 μM SNAP-Cell TMR Star (NEB) for 30 min. Nonreacted TMR Star was washed out, cells were fixed in 3.7% paraformaldehyde/0.3% Triton-X100 and nuclei were counterstained with DAPI.

    Article Title: Lipids Regulate Lck Protein Activity through Their Interactions with the Lck Src Homology 2 Domain
    Article Snippet: Simultaneous single molecule imaging of two proteins was performed in Jurkat cells using a total internal reflection fluorescence microscope as described previously ( ). .. JCaM1.6 Jurkat cells stably expressing Lck-GFP WT (or mutants) were transiently transfected with SNAP®-tagged TCR-ζ chain, which was subsequently labeled with SNAP-Cell® TMR-Star® (New England Biolabs). .. Labeled cells were washed, attached to a poly- l -lysine-coated glass plate for imaging, and stimulated with 10 μg/ml OKT3, and Lck and ζ molecules were simultaneously tracked.

    Article Title: Site-Specific Protein Labeling with SNAP-Tags
    Article Snippet: SNAP-tag fusion proteins labeled with SNAP-Surface Alexa Fluor 488 should have an excitation maximum at 496 nm and an emission maximum at 520 nm, and can be imaged with standard fluorescein filter sets. .. Cell permeable BG probe (e.g. SNAP-Cell TMR Star, NEB #S9105S).

    Article Title: Clathrin promotes centrosome integrity in early mitosis through stabilization of centrosomal ch-TOG
    Article Snippet: Precleared and IP samples were resolved by SDS-PAGE, transferred to nitrocellulose membranes, and analyzed by immunoblotting. .. For in vivo labeling of SNAP-tagged uLCa fusion proteins, live cells plated on coverslips were incubated (30 min at 37°C in 5% CO2 ) with 3 µM SNAP-TMR-Star (New England Biolabs, Inc.) diluted in normal growth media with serum. .. Cells were then washed three times with normal growth media with serum, incubated (37°C in 5% CO2 ) an additional 30 min, and then imaged using confocal laser-scanning microscopy, as detailed under Time-lapse confocal microscopy imaging.

    Article Title: Tyrosination of α‐tubulin controls the initiation of processive dynein–dynactin motility
    Article Snippet: The DDB complex was prepared by adding recombinant strepII‐SNAPf‐tagged BiCD2 (N‐terminal construct encompassing amino acids 25–400) to high‐speed porcine brain lysates as previously described (McKenney et al , ). .. The DDB complexes were fluorescently labeled with excess SNAP‐Cell TMR‐Star dye (NEB) during purification as described (McKenney et al , ), and aliquots of eluted DDB were flash‐frozen in LN2 and stored at −80°C. .. We note that freezing the complex leads to an apparently larger percentage of diffusive complexes in our assays (~15% for unfrozen versus ~30% for frozen).

    Article Title: The F-actin modifier villin regulates insulin granule dynamics and exocytosis downstream of islet cell autoantigen 512
    Article Snippet: 2.9 Prior to imaging, cells grown in an open chamber were incubated in resting media and transferred onto a thermostat-controlled (37 °C) stage. .. Insulin-SNAP+ SGs were labeled as previously described with the SNAP substrate TMR-Star (New England Biolabs, Ipswich, MA). .. The SGs were visualized in cells cotransfected with either EGFP or villin-GFP after treatment with the Ica512 , villin , or scramble siRNAs.

    Article Title: Newly produced synaptic vesicle proteins are preferentially used in synaptic transmission
    Article Snippet: Click labeling was performed after fixation with a commercial kit (Click‐iT Cell Reaction Buffer Kit; Invitrogen), and with 5 mM Chromeo494‐alkyne (Jena Bioscience, Jena, Germany). .. SNAP tag labeling was performed with the cell‐permeable dyes TMR‐Star and 647‐SiR (both from New England Biolabs, Ipswich, MA, USA). .. First, TMR‐Star was applied after 3–4 days of expression of the VAMP2‐TEV‐SNAP construct, for 15–30 min at 1 μM in the neurons’ own culture medium.

    SDS Page:

    Article Title: Molecular coordination of Staphylococcus aureus cell division
    Article Snippet: SNAP-Cell TMR-Star (New England Biolabs) was added to a 1 ml aliquot of mid-exponential phase (OD600 ~1) grown culture at a concentration of 500 nM and incubated at 37°C for 1 hr. .. Cells were washed three times by resuspension and centrifugation in PBS, resuspended in PBS supplemented with 200 μg ml−1 lysostaphin and 20 U ml−1 DNase I and lysed at 37°C for 30 min.

    Software:

    Article Title: A novel role for the histone acetyltransferase Hat1 in the CENP-A/CID assembly pathway in Drosophila melanogaster
    Article Snippet: After 48 h (Figure , Supplementary Figure S6) or 24 h (Supplementary Figure S6) chase, cells were pulse-labeled with 4.5 μM SNAP-Cell TMR Star (NEB) for 30 min. Nonreacted TMR Star was washed out, cells were fixed in 3.7% paraformaldehyde/0.3% Triton-X100 and nuclei were counterstained with DAPI. .. After 48 h (Figure , Supplementary Figure S6) or 24 h (Supplementary Figure S6) chase, cells were pulse-labeled with 4.5 μM SNAP-Cell TMR Star (NEB) for 30 min. Nonreacted TMR Star was washed out, cells were fixed in 3.7% paraformaldehyde/0.3% Triton-X100 and nuclei were counterstained with DAPI.

    Article Title: A Recombination Directionality Factor Controls the Cell Type-Specific Activation of σK and the Fidelity of Spore Development in Clostridium difficile
    Article Snippet: For SNAP staining, culture samples of 1 ml were stained for 30 min with 250 nM SNAP-Cell TMR-Star (New England Biolabs) as described before [ ]. .. For SNAP staining, culture samples of 1 ml were stained for 30 min with 250 nM SNAP-Cell TMR-Star (New England Biolabs) as described before [ ].

    Article Title: The F-actin modifier villin regulates insulin granule dynamics and exocytosis downstream of islet cell autoantigen 512
    Article Snippet: Insulin-SNAP+ SGs were labeled as previously described with the SNAP substrate TMR-Star (New England Biolabs, Ipswich, MA). .. Insulin-SNAP+ SGs were labeled as previously described with the SNAP substrate TMR-Star (New England Biolabs, Ipswich, MA).

    Binding Assay:

    Article Title: Tyrosination of α‐tubulin controls the initiation of processive dynein–dynactin motility
    Article Snippet: The protein was subjected to a cycle of MT binding and release by ATP to select for active motors. .. The DDB complexes were fluorescently labeled with excess SNAP‐Cell TMR‐Star dye (NEB) during purification as described (McKenney et al , ), and aliquots of eluted DDB were flash‐frozen in LN2 and stored at −80°C.

    Spectrophotometry:

    Article Title: Lissencephaly-1 is a context-dependent regulator of the human dynein complex
    Article Snippet: IgG Sepharose 6 Fast Flow beads bound to SNAPf ::dynein complexes or LIS1::SNAPf were incubated at 4°C for 40 min with ~5 μM SNAP-Surface Alexa Fluor 647 or SNAP-Cell TMR-Star (NEB) for SNAPf ::dynein, or ~40 μM SNAP-Cell TMR-Star for LIS1::SNAPf . .. IgG Sepharose 6 Fast Flow beads bound to SNAPf ::dynein complexes or LIS1::SNAPf were incubated at 4°C for 40 min with ~5 μM SNAP-Surface Alexa Fluor 647 or SNAP-Cell TMR-Star (NEB) for SNAPf ::dynein, or ~40 μM SNAP-Cell TMR-Star for LIS1::SNAPf .

    Concentration Assay:

    Article Title: Molecular coordination of Staphylococcus aureus cell division
    Article Snippet: The volume was then calculated based on a prolate spheroid shape with volume V = 4 3 π a b 2 , where a and b are the dimensions of the long and short axis respectively. .. SNAP-Cell TMR-Star (New England Biolabs) was added to a 1 ml aliquot of mid-exponential phase (OD600 ~1) grown culture at a concentration of 500 nM and incubated at 37°C for 1 hr. .. Cells were washed three times by resuspension and centrifugation in PBS, resuspended in PBS supplemented with 200 μg ml−1 lysostaphin and 20 U ml−1 DNase I and lysed at 37°C for 30 min.

    Construct:

    Article Title: Tyrosination of α‐tubulin controls the initiation of processive dynein–dynactin motility
    Article Snippet: Paragraph title: Purification of dynein–dynactin–BicD2 (DDB) complex and p150 constructs ... The DDB complexes were fluorescently labeled with excess SNAP‐Cell TMR‐Star dye (NEB) during purification as described (McKenney et al , ), and aliquots of eluted DDB were flash‐frozen in LN2 and stored at −80°C.

    Staining:

    Article Title: A SNAP-Tagged Derivative of HIV-1--A Versatile Tool to Study Virus-Cell Interactions
    Article Snippet: Paragraph title: Intracellular staining of Gag.SNAP ... Hela cells seeded on coverslips 24 h prior to the experiment were transfected with FuGene6 (Roche) and incubated at 37°C for 24 h. For SNAP-labelling cells were incubated with 1 µM SNAP-cell TMR-Star (New England Biolabs) in DMEM for 15 min at 37°C, washed and incubated in fresh DMEM for 45 min at 37°C to allow diffusion of excess substrate.

    Article Title: Comparison of a mouse and a novel human scFv-SNAP-auristatin F drug conjugate with potent activity against EGFR-overexpressing human solid tumor cells
    Article Snippet: Briefly, 7×103 cells were plated in a flat bottom black 96-well half-area microplate (µclear; Greiner Bio-One) in a total volume of 50 µL cell culture medium without G418. .. The recombinant tubulin structure was stained directly by adding 50 µL/well of a solution in growth medium containing 30 nM SNAP-Cell TMR-Star (NEB) for labeling the tubulin and 1 µg/mL 4′,6-diamidino-2-phenylindole (Thermo Fisher Scientific) for nuclei counterstaining. .. The cells were incubated overnight at 37°C, 5% CO2 and 100% humidity, before incubation with 50, 150, or 250 nM of the EGFR-specific ADCs.

    Article Title: Genome-Wide Analysis of Cell Type-Specific Gene Transcription during Spore Formation in Clostridium difficileThe Spore Differentiation Pathway in the Enteric Pathogen Clostridium difficile
    Article Snippet: The cells were washed with 1 ml of PBS and ressuspended in 0.1 ml of PBS supplemented with the membrane dye FM4-64 (10 µg.ml−1 ) and the DNA stain DAPI (4′,6-diamidino-2-phenylindole; 50 µg.ml−1 ) (Molecular Probes, Invitrogen). .. For SNAP staining, 1 ml samples were stained for 30 min with 50 nM SNAP-Cell TMR-Star (New England Biolabs) as described . .. Cells were washed four times by centrifugation (4000 g , 5 min) and ressupended in 1 ml of PBS.

    Article Title: A Recombination Directionality Factor Controls the Cell Type-Specific Activation of σK and the Fidelity of Spore Development in Clostridium difficile
    Article Snippet: The cells were washed with 1ml of PBS and resuspended in 0.1 ml of PBS supplemented with the membrane dye Mitotracker Green (MTG) at 0.5 μg.ml-1 and the DNA stain DAPI (4',6-diamidino-2-phenylindole; 50 μg.ml-1 ) (Invitrogen). .. For SNAP staining, culture samples of 1 ml were stained for 30 min with 250 nM SNAP-Cell TMR-Star (New England Biolabs) as described before [ ]. .. Cells were washed four times by centrifugation (4000 g , 5 min) and ressupended in 1ml of PBS.

    High Content Screening:

    Article Title: Comparison of a mouse and a novel human scFv-SNAP-auristatin F drug conjugate with potent activity against EGFR-overexpressing human solid tumor cells
    Article Snippet: The recombinant tubulin structure was stained directly by adding 50 µL/well of a solution in growth medium containing 30 nM SNAP-Cell TMR-Star (NEB) for labeling the tubulin and 1 µg/mL 4′,6-diamidino-2-phenylindole (Thermo Fisher Scientific) for nuclei counterstaining. .. The cells were incubated overnight at 37°C, 5% CO2 and 100% humidity, before incubation with 50, 150, or 250 nM of the EGFR-specific ADCs.

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    New England Biolabs fluorescent snap substrate
    Fluorescent Snap Substrate, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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