bg549 surface  (New England Biolabs)


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  • 95
    Name:
    SNAP Surface 549
    Description:
    SNAP Surface 549 50 nmol
    Catalog Number:
    s9112s
    Price:
    344
    Size:
    50 nmol
    Category:
    Fluorochromes
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    Structured Review

    New England Biolabs bg549 surface
    SNAP Surface 549
    SNAP Surface 549 50 nmol
    https://www.bioz.com/result/bg549 surface/product/New England Biolabs
    Average 95 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    bg549 surface - by Bioz Stars, 2020-02
    95/100 stars

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    Related Articles

    Electrophoresis:

    Article Title: A Single Mechanism of Biogenesis, Initiated and Directed by PIWI Proteins, Explains piRNA Production in Most Animals
    Article Snippet: .. SNAP-MIWI was labeled with SNAP substrate SNAP-Surface 549 (NEB, S9112) and resolved by electrophoresis through a 4–20% gradient SDS- polyacrylamide gel (Bio-Rad Laboratories, 5671085). .. The concentration of full-length SNAP-MIWI was determined by comparison to a standard curve of purified, SNAP- Surface 549 labeled SNAP protein (Typhoon FLA 7000; GE Lifesciences).

    Incubation:

    Article Title: Alternative transcription cycle for bacterial RNA polymerase
    Article Snippet: .. RNAP-SNAP was labeled with the DY-549 dye, yielding RNAP549 , as follows: 20 μL of 15 μM RNAP-SNAP was dialyzed into 3 L of labeling buffer (10 mM Tris-HCl, pH 8.0, 40 mM KCl, 5 mM MgCl2 , 20 μM ZnCl2 , and 1 mM dithiothreitol (DTT)) at 4 °C for 4 h. The resulting product (typically 50–100 μL of 5–20 μM of protein) was mixed with an equimolar amount of SNAP-Surface 549 (New England Biolabs; 1 mM in DMSO) and incubated at room temperature for 30 min, then mixed with an equal volume of labeling buffer supplemented with 60% glycerol to yield RNAP549 in reconstitution buffer (10 mM Tris-HCl, pH 8.0, 30% glycerol, 0.1 mM EDTA, 100 mM NaCl, 20 mM KCl, 20 μM ZnCl2 , 3 mM MgCl2 , and 0.6 mM DTT). ..

    Article Title: Nerve growth factor–mediated photoablation of nociceptors reduces pain behavior in mice
    Article Snippet: For the labelling, 1-µM NGFR121W-SNAP was coupled with 3-µM BG549 surface (NEB # S9112) for 1 hour at 37°C in calcium imaging buffer (NaCl 140 mM; KCl 4 mM; CaCl2 2 mM; MgCl2 1 mM; NaOH; 4.55 mM; glucose 5 mM; and HEPES 10 mM; pH 7.4). .. Cells were incubated with the coupling reaction for 15 to 20 minutes at 37°C, then washed 3 times in calcium imaging buffer.

    Article Title: Synergistic assembly of human pre-spliceosomes across introns and exons
    Article Snippet: .. After centrifuging the lysate for 20 s at 12,000 × g , the nuclear pellet was resuspended in 450 µl Buffer C [20 mM Tris, 25% (v/v) glycerol, 0.42 M NaCl, 1.5 mM MgCl2 , 0.2 mM EDTA and 0.5 mM DTT, pH 7.9 at 4°C, supplemented with complete protease inhibitor cocktail (Roche, 04693159001)] and rapidly stirred in a 2 ml round-bottom microcentrifuge tube with a 12.7 × 3 mm stir bar for 30 min. After clarifying the lysate by centrifuging for 10 min at 12,000 × g , the SNAP-Surface 549 dye-benzylguanine conjugate (New England BioLabs, S9112S) was added to the supernatant at a final concentration of 200 nM and incubated for 30 min at 30°C. .. After labeling, the supernatant was dialyzed 2 times for 2 hr each against Buffer E (20 mM Tris, 20% (v/v) glycerol, 0.1 M KCl, 0.2 mM EDTA and 0.5 mM DTT, pH 7.9 at 4°C).

    Article Title: A ligand-based system for receptor-specific delivery of proteins
    Article Snippet: In-vitro labelling For keratinocyte labelling, IL-31K138A SNAP, NGFR121W SNAP or SNAP were coupled with an excess of BG-Surface549 at a 1:1.5 molar ratio (NEB # S9112) for 2 hours at 37 °C in PBS (pH 7.4) or CIB buffer (NaCl 140 mM; KCl 4 mM; CaCl2 2 mM; MgCl2 1 mM; NaOH 4.55 mM; Glucose 5 mM; HEPES 10 mM; pH 7.4). .. Cells from wild type mice were incubated with the coupling reaction for 2 hours at 37 °C, then washed 3 times in PBS.

    Article Title: An engineered opsin monomer scrambles phospholipids
    Article Snippet: To the Neutravidin coated chambers, 10–20 nM of biotinylated monoclonal anti-FLAG antibody (Sigma, cat. no. F9291) was added and incubated for 30 mins at RT followed by washing with T50 buffer. .. The flow chamber was placed on an inverted TIRF microscope (Olympus IX73 with cellTIRF system) and purified SNAP-surface549 (New England BioLabs) labeled QUAD-FLAG-SNAP or WT-FLAG-SNAP was immobilized at a density that allowed clear resolution of individual spots.

    Expressing:

    Article Title: Synergistic assembly of human pre-spliceosomes across introns and exons
    Article Snippet: Expression levels of the fSNAP fusion proteins were adjusted to endogenous level by inducing the U1-70K-fSNAP, U2B''-fSNAP, and Snu114-fSNAP Flp-In cell lines with 6 ng/ml, 3 ng/ml, and 3 ng/ml Doxycycline (BD Biosciences, 631311), respectively. .. After centrifuging the lysate for 20 s at 12,000 × g , the nuclear pellet was resuspended in 450 µl Buffer C [20 mM Tris, 25% (v/v) glycerol, 0.42 M NaCl, 1.5 mM MgCl2 , 0.2 mM EDTA and 0.5 mM DTT, pH 7.9 at 4°C, supplemented with complete protease inhibitor cocktail (Roche, 04693159001)] and rapidly stirred in a 2 ml round-bottom microcentrifuge tube with a 12.7 × 3 mm stir bar for 30 min. After clarifying the lysate by centrifuging for 10 min at 12,000 × g , the SNAP-Surface 549 dye-benzylguanine conjugate (New England BioLabs, S9112S) was added to the supernatant at a final concentration of 200 nM and incubated for 30 min at 30°C.

    BIA-KA:

    Article Title: A Single Mechanism of Biogenesis, Initiated and Directed by PIWI Proteins, Explains piRNA Production in Most Animals
    Article Snippet: SNAP-MIWI was labeled with SNAP substrate SNAP-Surface 549 (NEB, S9112) and resolved by electrophoresis through a 4–20% gradient SDS- polyacrylamide gel (Bio-Rad Laboratories, 5671085). .. The concentration of purified SNAP protein (gift from Moore Lab) was determined before labeling by BCA assay and by measuring its absorbance at 280 nm (ε = 20970 M-1 cm-1 in water assuming all Cys residues are reduced).

    Western Blot:

    Article Title: A novel two-step genome editing strategy with CRISPR-Cas9 provides new insights into telomerase action and TERT gene expression
    Article Snippet: Paragraph title: Western blot and SNAP-tag fluorescence labeling ... To detect the SNAP-tag, 10 μM SNAP Surface® 594 (New England Biolabs, S9112S) was added to the input samples at the beginning of the IP.

    Transfection:

    Article Title: Molecular details of dimerization kinetics reveal negligible populations of transient µ-opioid receptor homodimers at physiological concentrations
    Article Snippet: .. 24–36 hours after transfection cells were labeled with SNAP-Surface Dyes 549 and Alexa Fluor 647 (New England Biolabs, Frankfurt am Main, Germany) according to the manufacturer’s instructions. ..

    Article Title: PD-L1:CD80 Cis-Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways
    Article Snippet: .. 24 h post transfection, cell was stained with a 1:1 mixture of SS549 (energy donor, NEB S9112S) and SSAF647 (energy acceptor) at 37°C, 5% CO2 for 30 min, and washed 3 times with 1x PBS (pH 7.4). .. For the atezolizumab treated condition, 20 μg/mL atezolizumab was included in the staining solution.

    Article Title: Nerve growth factor–mediated photoablation of nociceptors reduces pain behavior in mice
    Article Snippet: In vitro experiments Hek293T cells were transfected with TrkA, TrkB, TrkC, and p75 plasmid using Lipofectamine 2000 (Thermo). .. For the labelling, 1-µM NGFR121W-SNAP was coupled with 3-µM BG549 surface (NEB # S9112) for 1 hour at 37°C in calcium imaging buffer (NaCl 140 mM; KCl 4 mM; CaCl2 2 mM; MgCl2 1 mM; NaOH; 4.55 mM; glucose 5 mM; and HEPES 10 mM; pH 7.4).

    Protease Inhibitor:

    Article Title: Synergistic assembly of human pre-spliceosomes across introns and exons
    Article Snippet: .. After centrifuging the lysate for 20 s at 12,000 × g , the nuclear pellet was resuspended in 450 µl Buffer C [20 mM Tris, 25% (v/v) glycerol, 0.42 M NaCl, 1.5 mM MgCl2 , 0.2 mM EDTA and 0.5 mM DTT, pH 7.9 at 4°C, supplemented with complete protease inhibitor cocktail (Roche, 04693159001)] and rapidly stirred in a 2 ml round-bottom microcentrifuge tube with a 12.7 × 3 mm stir bar for 30 min. After clarifying the lysate by centrifuging for 10 min at 12,000 × g , the SNAP-Surface 549 dye-benzylguanine conjugate (New England BioLabs, S9112S) was added to the supernatant at a final concentration of 200 nM and incubated for 30 min at 30°C. .. After labeling, the supernatant was dialyzed 2 times for 2 hr each against Buffer E (20 mM Tris, 20% (v/v) glycerol, 0.1 M KCl, 0.2 mM EDTA and 0.5 mM DTT, pH 7.9 at 4°C).

    Cell Culture:

    Article Title: Nerve growth factor–mediated photoablation of nociceptors reduces pain behavior in mice
    Article Snippet: For the labelling, 1-µM NGFR121W-SNAP was coupled with 3-µM BG549 surface (NEB # S9112) for 1 hour at 37°C in calcium imaging buffer (NaCl 140 mM; KCl 4 mM; CaCl2 2 mM; MgCl2 1 mM; NaOH; 4.55 mM; glucose 5 mM; and HEPES 10 mM; pH 7.4). .. To assess NGF signaling, PC12 cells were cultured on collagen IV–coated 6-well plate in DMEM/F12 medium containing 1% horse serum.

    Article Title: Agonist-induced dimer dissociation as a macromolecular step in G protein-coupled receptor signaling
    Article Snippet: Kinetic determination of mobile fractions of crosslinked V5-FZD6 -mCherry and FZD6 -GFP with WNT stimulation over time were performed in one cell culture well. .. The cell-impermeable SNAP substrate SNAP-Surface 549 (New England Biolabs) was added (1:1000, 15 min), washed twice in BE buffer and imaged using the 561 nm laser line.

    other:

    Article Title: Dynamic caveolae exclude bulk membrane proteins and are required for sorting of excess glycosphingolipids
    Article Snippet: The reagents used in this study are: cholera toxin B subunit conjugated to HRP (Molecular Probes, C3478) or alexa fluor 555 (Molecular Probes, C34776) or alexa fluor 647 (Molecular Probes, C34778); alexa fluor 647-conjugated human transferrin (Invitrogen, T23366); SNAP Cell surface Reagent 549 (New England Biolabs, S9112S); cyclohexamide (Santa-Cruz, sc-3508); okadaic acid (Santa-Cruz, sc-95060);and Magic Red Cathepsin L kit (MR-FR2, Immunochemistry Technologies).

    Imaging:

    Article Title: Molecular details of dimerization kinetics reveal negligible populations of transient µ-opioid receptor homodimers at physiological concentrations
    Article Snippet: 24–36 hours after transfection cells were labeled with SNAP-Surface Dyes 549 and Alexa Fluor 647 (New England Biolabs, Frankfurt am Main, Germany) according to the manufacturer’s instructions. .. After incubation cells were washed three times with DMEM and immediately taken for imaging in HBSS-Buffer.

    Article Title: Nerve growth factor–mediated photoablation of nociceptors reduces pain behavior in mice
    Article Snippet: .. For the labelling, 1-µM NGFR121W-SNAP was coupled with 3-µM BG549 surface (NEB # S9112) for 1 hour at 37°C in calcium imaging buffer (NaCl 140 mM; KCl 4 mM; CaCl2 2 mM; MgCl2 1 mM; NaOH; 4.55 mM; glucose 5 mM; and HEPES 10 mM; pH 7.4). .. The coupling reaction was filtered through a PD MiniTrap G-25 column (GE Healthcare #28-9180-07) to remove the excess of BG.

    Article Title: A novel two-step genome editing strategy with CRISPR-Cas9 provides new insights into telomerase action and TERT gene expression
    Article Snippet: The signals were detected with a FluorChem HD2 imaging system (Alpha Innotech) and quantified with ImageQuant TL v2005 software. .. To detect the SNAP-tag, 10 μM SNAP Surface® 594 (New England Biolabs, S9112S) was added to the input samples at the beginning of the IP.

    Article Title: Agonist-induced dimer dissociation as a macromolecular step in G protein-coupled receptor signaling
    Article Snippet: Paragraph title: FRAP and cellular imaging ... The cell-impermeable SNAP substrate SNAP-Surface 549 (New England Biolabs) was added (1:1000, 15 min), washed twice in BE buffer and imaged using the 561 nm laser line.

    Protein Concentration:

    Article Title: Comprehensive and quantitative mapping of RNA–protein interactions across a transcribed eukaryotic genome
    Article Snippet: Protein concentration was measured both by Bradford dye reagent and A 280 absorbance. .. Subsequently, Vts1(442–523)-SNAP-His6 was fluorescently labeled using SNAP-Surface549 (NEB).

    Staining:

    Article Title: PD-L1:CD80 Cis-Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways
    Article Snippet: .. 24 h post transfection, cell was stained with a 1:1 mixture of SS549 (energy donor, NEB S9112S) and SSAF647 (energy acceptor) at 37°C, 5% CO2 for 30 min, and washed 3 times with 1x PBS (pH 7.4). .. For the atezolizumab treated condition, 20 μg/mL atezolizumab was included in the staining solution.

    In Vivo:

    Article Title: Single-molecule Studies of Origin Licensing Reveal Mechanisms Ensuring Bidirectional Helicase Loading
    Article Snippet: The Ubiquitin ( in vivo ) and GST-SUMO (using Ulp1 protease) fusions were removed to reveal three N-terminal glycines required for sortase labeling. .. SNAP-Surface549 (NEB, SNAP549 in the manuscript) or SNAP-Janelia Fluor 646 (SNAPJF646; ) was coupled to SNAP-tagged Mcm2-7 (See for these purification protocols).

    Fluorescence:

    Article Title: A novel two-step genome editing strategy with CRISPR-Cas9 provides new insights into telomerase action and TERT gene expression
    Article Snippet: Paragraph title: Western blot and SNAP-tag fluorescence labeling ... To detect the SNAP-tag, 10 μM SNAP Surface® 594 (New England Biolabs, S9112S) was added to the input samples at the beginning of the IP.

    Article Title: Agonist-induced dimer dissociation as a macromolecular step in G protein-coupled receptor signaling
    Article Snippet: The mobile fraction was defined by the average of the fluorescence recovery assessed between time 85–101 s (including pre-bleach measurements). .. The cell-impermeable SNAP substrate SNAP-Surface 549 (New England Biolabs) was added (1:1000, 15 min), washed twice in BE buffer and imaged using the 561 nm laser line.

    Mutagenesis:

    Article Title: PD-L1:CD80 Cis-Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways
    Article Snippet: For , SNAP-tagged full length CD80 (SNAP–CD80), CD80 mutant (SNAP–CD80 I92R) or CD86 (SNAP–CD86) was transfected to either HEK293T cells or CLIP–PD-L1 transduced HEK293T cells using polyethylenimine. .. 24 h post transfection, cell was stained with a 1:1 mixture of SS549 (energy donor, NEB S9112S) and SSAF647 (energy acceptor) at 37°C, 5% CO2 for 30 min, and washed 3 times with 1x PBS (pH 7.4).

    Flow Cytometry:

    Article Title: Comprehensive and quantitative mapping of RNA–protein interactions across a transcribed eukaryotic genome
    Article Snippet: The dialysate was then mixed for 1 h with 2 mL of a 50% (vol/vol) slurry of SP-Sepharose resin (GE) that had been equilibrated in buffer C. The resin (1 mL) was then poured into a column and Vts1(442–523)-SNAP-His6 was recovered as a flow-through and subsequently concentrated by centrifugal ultrafiltration ( ). .. Subsequently, Vts1(442–523)-SNAP-His6 was fluorescently labeled using SNAP-Surface549 (NEB).

    Article Title: An engineered opsin monomer scrambles phospholipids
    Article Snippet: .. The flow chamber was placed on an inverted TIRF microscope (Olympus IX73 with cellTIRF system) and purified SNAP-surface549 (New England BioLabs) labeled QUAD-FLAG-SNAP or WT-FLAG-SNAP was immobilized at a density that allowed clear resolution of individual spots. ..

    Microscopy:

    Article Title: Molecular details of dimerization kinetics reveal negligible populations of transient µ-opioid receptor homodimers at physiological concentrations
    Article Snippet: Paragraph title: Microscope sample preparation and SNAP-labeling ... 24–36 hours after transfection cells were labeled with SNAP-Surface Dyes 549 and Alexa Fluor 647 (New England Biolabs, Frankfurt am Main, Germany) according to the manufacturer’s instructions.

    Article Title: PD-L1:CD80 Cis-Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways
    Article Snippet: Images were acquired with an Olympus FV1000 confocal microscope by exciting CS547 (energy donor) at 543 nm and SSAF647 (energy acceptor) at 635 nm. .. 24 h post transfection, cell was stained with a 1:1 mixture of SS549 (energy donor, NEB S9112S) and SSAF647 (energy acceptor) at 37°C, 5% CO2 for 30 min, and washed 3 times with 1x PBS (pH 7.4).

    Article Title: An engineered opsin monomer scrambles phospholipids
    Article Snippet: .. The flow chamber was placed on an inverted TIRF microscope (Olympus IX73 with cellTIRF system) and purified SNAP-surface549 (New England BioLabs) labeled QUAD-FLAG-SNAP or WT-FLAG-SNAP was immobilized at a density that allowed clear resolution of individual spots. ..

    Purification:

    Article Title: Alternative transcription cycle for bacterial RNA polymerase
    Article Snippet: Proteins E. coli core RNAP (αββ′ω) with a SNAP tag on the c-terminus of β´ (RNAP-SNAP) and wild-type σ70 protein were expressed and purified . .. RNAP-SNAP was labeled with the DY-549 dye, yielding RNAP549 , as follows: 20 μL of 15 μM RNAP-SNAP was dialyzed into 3 L of labeling buffer (10 mM Tris-HCl, pH 8.0, 40 mM KCl, 5 mM MgCl2 , 20 μM ZnCl2 , and 1 mM dithiothreitol (DTT)) at 4 °C for 4 h. The resulting product (typically 50–100 μL of 5–20 μM of protein) was mixed with an equimolar amount of SNAP-Surface 549 (New England Biolabs; 1 mM in DMSO) and incubated at room temperature for 30 min, then mixed with an equal volume of labeling buffer supplemented with 60% glycerol to yield RNAP549 in reconstitution buffer (10 mM Tris-HCl, pH 8.0, 30% glycerol, 0.1 mM EDTA, 100 mM NaCl, 20 mM KCl, 20 μM ZnCl2 , 3 mM MgCl2 , and 0.6 mM DTT).

    Article Title: A Single Mechanism of Biogenesis, Initiated and Directed by PIWI Proteins, Explains piRNA Production in Most Animals
    Article Snippet: SNAP-MIWI was labeled with SNAP substrate SNAP-Surface 549 (NEB, S9112) and resolved by electrophoresis through a 4–20% gradient SDS- polyacrylamide gel (Bio-Rad Laboratories, 5671085). .. The concentration of full-length SNAP-MIWI was determined by comparison to a standard curve of purified, SNAP- Surface 549 labeled SNAP protein (Typhoon FLA 7000; GE Lifesciences).

    Article Title: Single-molecule Studies of Origin Licensing Reveal Mechanisms Ensuring Bidirectional Helicase Loading
    Article Snippet: .. SNAP-Surface549 (NEB, SNAP549 in the manuscript) or SNAP-Janelia Fluor 646 (SNAPJF646; ) was coupled to SNAP-tagged Mcm2-7 (See for these purification protocols). .. For Sortase labeling, peptide-coupled proteins were separated from uncoupled proteins using Complete-His-Tag Resin (Roche).

    Article Title: An engineered opsin monomer scrambles phospholipids
    Article Snippet: .. The flow chamber was placed on an inverted TIRF microscope (Olympus IX73 with cellTIRF system) and purified SNAP-surface549 (New England BioLabs) labeled QUAD-FLAG-SNAP or WT-FLAG-SNAP was immobilized at a density that allowed clear resolution of individual spots. ..

    Article Title: Quantitative analysis of RNA-protein interactions on a massively parallel array for mapping biophysical and evolutionary landscapes
    Article Snippet: .. Labeling MS2 Coat Protein with SNAPtag Substrate 5 μM purified SNAPtag-MS2 was labeled with SNAP-Surface 549 fluor (NEB) at 37°C for 30 minutes in 50 mM Tris pH 8.0, 100 mM NaCl, 0.1% Tween 20, 1 mM DTT, and 10 μM SNAPSurface 549. .. Excess SNAP-Surface 549 was removed using Zeba Spin Desalting Columns (Thermo) equilibrated with TMK Buffer (100 mM Tris-HCl pH 8.0, 80 mM KCl, 10 mM MgCl2 , 1 mM DTT).

    Protein Purification:

    Article Title: Comprehensive and quantitative mapping of RNA–protein interactions across a transcribed eukaryotic genome
    Article Snippet: Paragraph title: Vts1 Protein Purification, Labeling, and Quantification. ... Subsequently, Vts1(442–523)-SNAP-His6 was fluorescently labeled using SNAP-Surface549 (NEB).

    Article Title: Single-molecule Studies of Origin Licensing Reveal Mechanisms Ensuring Bidirectional Helicase Loading
    Article Snippet: Paragraph title: Protein Purification and Labeling ... SNAP-Surface549 (NEB, SNAP549 in the manuscript) or SNAP-Janelia Fluor 646 (SNAPJF646; ) was coupled to SNAP-tagged Mcm2-7 (See for these purification protocols).

    Labeling:

    Article Title: Molecular details of dimerization kinetics reveal negligible populations of transient µ-opioid receptor homodimers at physiological concentrations
    Article Snippet: .. 24–36 hours after transfection cells were labeled with SNAP-Surface Dyes 549 and Alexa Fluor 647 (New England Biolabs, Frankfurt am Main, Germany) according to the manufacturer’s instructions. ..

    Article Title: Alternative transcription cycle for bacterial RNA polymerase
    Article Snippet: .. RNAP-SNAP was labeled with the DY-549 dye, yielding RNAP549 , as follows: 20 μL of 15 μM RNAP-SNAP was dialyzed into 3 L of labeling buffer (10 mM Tris-HCl, pH 8.0, 40 mM KCl, 5 mM MgCl2 , 20 μM ZnCl2 , and 1 mM dithiothreitol (DTT)) at 4 °C for 4 h. The resulting product (typically 50–100 μL of 5–20 μM of protein) was mixed with an equimolar amount of SNAP-Surface 549 (New England Biolabs; 1 mM in DMSO) and incubated at room temperature for 30 min, then mixed with an equal volume of labeling buffer supplemented with 60% glycerol to yield RNAP549 in reconstitution buffer (10 mM Tris-HCl, pH 8.0, 30% glycerol, 0.1 mM EDTA, 100 mM NaCl, 20 mM KCl, 20 μM ZnCl2 , 3 mM MgCl2 , and 0.6 mM DTT). ..

    Article Title: PD-L1:CD80 Cis-Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways
    Article Snippet: Labeled cells were then fixed with 4% paraformaldehyde (PFA, Fisher Scientific, 50980494) and used for the FRET assay. .. 24 h post transfection, cell was stained with a 1:1 mixture of SS549 (energy donor, NEB S9112S) and SSAF647 (energy acceptor) at 37°C, 5% CO2 for 30 min, and washed 3 times with 1x PBS (pH 7.4).

    Article Title: Comprehensive and quantitative mapping of RNA–protein interactions across a transcribed eukaryotic genome
    Article Snippet: .. Subsequently, Vts1(442–523)-SNAP-His6 was fluorescently labeled using SNAP-Surface549 (NEB). .. Covalent labeling of the SNAP tag was conducted by incubating 80 µL of reaction mixture containing 50 mM Tris⋅HCl, pH 7.4, 100 mM NaCl, 0.1% Tween 20, 2 mM DTT, 10 μM Vts1(442–523)-SNAP-His6 , and 20 μM SNAP-Surface 549 (NEB) at 4 °C for 16 h. Excess fluorescent dye was removed using 7K MWCO Zeba spin desalting columns (Thermo) following manufacturer’s instructions, and the labeled protein was recovered in buffer TMK (100 mM Tris⋅HCl, pH 7.4, 80 mM KCl, 10 mM MgCl2 , 1 mM DTT) ( ).

    Article Title: A novel two-step genome editing strategy with CRISPR-Cas9 provides new insights into telomerase action and TERT gene expression
    Article Snippet: Paragraph title: Western blot and SNAP-tag fluorescence labeling ... To detect the SNAP-tag, 10 μM SNAP Surface® 594 (New England Biolabs, S9112S) was added to the input samples at the beginning of the IP.

    Article Title: Ribonucleoprotein purification and characterization using RNA Mango
    Article Snippet: Paragraph title: SNAP labeling of yeast whole-cell extract ... Yeast whole-cell extracts (500 µL) were first fluorescently tagged with the SNAP-Surface 549 tag (NEB) by incubating the extracts with 1 U/µL Murine RNase inhibitor (NEB), Protease Inhibitors (cOmplete mini at the recommended concentration, EDTA Free, Sigma), 5 mM DTT together with 5 µM SNAP-Surface 549 tag in Buffer B at RT for 30 min.

    Article Title: A Single Mechanism of Biogenesis, Initiated and Directed by PIWI Proteins, Explains piRNA Production in Most Animals
    Article Snippet: .. SNAP-MIWI was labeled with SNAP substrate SNAP-Surface 549 (NEB, S9112) and resolved by electrophoresis through a 4–20% gradient SDS- polyacrylamide gel (Bio-Rad Laboratories, 5671085). .. The concentration of full-length SNAP-MIWI was determined by comparison to a standard curve of purified, SNAP- Surface 549 labeled SNAP protein (Typhoon FLA 7000; GE Lifesciences).

    Article Title: Synergistic assembly of human pre-spliceosomes across introns and exons
    Article Snippet: After centrifuging the lysate for 20 s at 12,000 × g , the nuclear pellet was resuspended in 450 µl Buffer C [20 mM Tris, 25% (v/v) glycerol, 0.42 M NaCl, 1.5 mM MgCl2 , 0.2 mM EDTA and 0.5 mM DTT, pH 7.9 at 4°C, supplemented with complete protease inhibitor cocktail (Roche, 04693159001)] and rapidly stirred in a 2 ml round-bottom microcentrifuge tube with a 12.7 × 3 mm stir bar for 30 min. After clarifying the lysate by centrifuging for 10 min at 12,000 × g , the SNAP-Surface 549 dye-benzylguanine conjugate (New England BioLabs, S9112S) was added to the supernatant at a final concentration of 200 nM and incubated for 30 min at 30°C. .. After labeling, the supernatant was dialyzed 2 times for 2 hr each against Buffer E (20 mM Tris, 20% (v/v) glycerol, 0.1 M KCl, 0.2 mM EDTA and 0.5 mM DTT, pH 7.9 at 4°C).

    Article Title: Single-molecule Studies of Origin Licensing Reveal Mechanisms Ensuring Bidirectional Helicase Loading
    Article Snippet: Paragraph title: Protein Purification and Labeling ... SNAP-Surface549 (NEB, SNAP549 in the manuscript) or SNAP-Janelia Fluor 646 (SNAPJF646; ) was coupled to SNAP-tagged Mcm2-7 (See for these purification protocols).

    Article Title: An engineered opsin monomer scrambles phospholipids
    Article Snippet: .. The flow chamber was placed on an inverted TIRF microscope (Olympus IX73 with cellTIRF system) and purified SNAP-surface549 (New England BioLabs) labeled QUAD-FLAG-SNAP or WT-FLAG-SNAP was immobilized at a density that allowed clear resolution of individual spots. ..

    Article Title: Quantitative analysis of RNA-protein interactions on a massively parallel array for mapping biophysical and evolutionary landscapes
    Article Snippet: .. Labeling MS2 Coat Protein with SNAPtag Substrate 5 μM purified SNAPtag-MS2 was labeled with SNAP-Surface 549 fluor (NEB) at 37°C for 30 minutes in 50 mM Tris pH 8.0, 100 mM NaCl, 0.1% Tween 20, 1 mM DTT, and 10 μM SNAPSurface 549. .. Excess SNAP-Surface 549 was removed using Zeba Spin Desalting Columns (Thermo) equilibrated with TMK Buffer (100 mM Tris-HCl pH 8.0, 80 mM KCl, 10 mM MgCl2 , 1 mM DTT).

    Confocal Microscopy:

    Article Title: PD-L1:CD80 Cis-Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways
    Article Snippet: Paragraph title: Confocal Microscopy Based FRET Assay with HEK293T Cells ... 24 h post transfection, cell was stained with a 1:1 mixture of SS549 (energy donor, NEB S9112S) and SSAF647 (energy acceptor) at 37°C, 5% CO2 for 30 min, and washed 3 times with 1x PBS (pH 7.4).

    FACS:

    Article Title: A Single Mechanism of Biogenesis, Initiated and Directed by PIWI Proteins, Explains piRNA Production in Most Animals
    Article Snippet: SNAP-MIWI was labeled with SNAP substrate SNAP-Surface 549 (NEB, S9112) and resolved by electrophoresis through a 4–20% gradient SDS- polyacrylamide gel (Bio-Rad Laboratories, 5671085). .. SNAP-MIWI of known concentration was then used to estimate the number of MIWI molecules present in FACS-purified germ cells from mouse testes by quantitative immunoblotting using anti-MIWI antibody.

    Mouse Assay:

    Article Title: A ligand-based system for receptor-specific delivery of proteins
    Article Snippet: In-vitro labelling For keratinocyte labelling, IL-31K138A SNAP, NGFR121W SNAP or SNAP were coupled with an excess of BG-Surface549 at a 1:1.5 molar ratio (NEB # S9112) for 2 hours at 37 °C in PBS (pH 7.4) or CIB buffer (NaCl 140 mM; KCl 4 mM; CaCl2 2 mM; MgCl2 1 mM; NaOH 4.55 mM; Glucose 5 mM; HEPES 10 mM; pH 7.4). .. Cells from wild type mice were incubated with the coupling reaction for 2 hours at 37 °C, then washed 3 times in PBS.

    SDS Page:

    Article Title: Comprehensive and quantitative mapping of RNA–protein interactions across a transcribed eukaryotic genome
    Article Snippet: The elution profile was monitored by SDS/PAGE. .. Subsequently, Vts1(442–523)-SNAP-His6 was fluorescently labeled using SNAP-Surface549 (NEB).

    Plasmid Preparation:

    Article Title: Nerve growth factor–mediated photoablation of nociceptors reduces pain behavior in mice
    Article Snippet: In vitro experiments Hek293T cells were transfected with TrkA, TrkB, TrkC, and p75 plasmid using Lipofectamine 2000 (Thermo). .. For the labelling, 1-µM NGFR121W-SNAP was coupled with 3-µM BG549 surface (NEB # S9112) for 1 hour at 37°C in calcium imaging buffer (NaCl 140 mM; KCl 4 mM; CaCl2 2 mM; MgCl2 1 mM; NaOH; 4.55 mM; glucose 5 mM; and HEPES 10 mM; pH 7.4).

    Software:

    Article Title: A novel two-step genome editing strategy with CRISPR-Cas9 provides new insights into telomerase action and TERT gene expression
    Article Snippet: The signals were detected with a FluorChem HD2 imaging system (Alpha Innotech) and quantified with ImageQuant TL v2005 software. .. To detect the SNAP-tag, 10 μM SNAP Surface® 594 (New England Biolabs, S9112S) was added to the input samples at the beginning of the IP.

    Article Title: An engineered opsin monomer scrambles phospholipids
    Article Snippet: The flow chamber was placed on an inverted TIRF microscope (Olympus IX73 with cellTIRF system) and purified SNAP-surface549 (New England BioLabs) labeled QUAD-FLAG-SNAP or WT-FLAG-SNAP was immobilized at a density that allowed clear resolution of individual spots. .. Images were acquired with a scMOS detector camera (Hamamatsu ORCA-flash4.0 V3) at 20 Hz using Olympus cellSens software.

    Sample Prep:

    Article Title: Molecular details of dimerization kinetics reveal negligible populations of transient µ-opioid receptor homodimers at physiological concentrations
    Article Snippet: Paragraph title: Microscope sample preparation and SNAP-labeling ... 24–36 hours after transfection cells were labeled with SNAP-Surface Dyes 549 and Alexa Fluor 647 (New England Biolabs, Frankfurt am Main, Germany) according to the manufacturer’s instructions.

    In Vitro:

    Article Title: Nerve growth factor–mediated photoablation of nociceptors reduces pain behavior in mice
    Article Snippet: Paragraph title: 3.4. In vitro experiments ... For the labelling, 1-µM NGFR121W-SNAP was coupled with 3-µM BG549 surface (NEB # S9112) for 1 hour at 37°C in calcium imaging buffer (NaCl 140 mM; KCl 4 mM; CaCl2 2 mM; MgCl2 1 mM; NaOH; 4.55 mM; glucose 5 mM; and HEPES 10 mM; pH 7.4).

    Article Title: A ligand-based system for receptor-specific delivery of proteins
    Article Snippet: .. In-vitro labelling For keratinocyte labelling, IL-31K138A SNAP, NGFR121W SNAP or SNAP were coupled with an excess of BG-Surface549 at a 1:1.5 molar ratio (NEB # S9112) for 2 hours at 37 °C in PBS (pH 7.4) or CIB buffer (NaCl 140 mM; KCl 4 mM; CaCl2 2 mM; MgCl2 1 mM; NaOH 4.55 mM; Glucose 5 mM; HEPES 10 mM; pH 7.4). .. The coupling reaction was passed through a PD MiniTrap G-25 column (GE Healthcare #28-9180-07) to remove the excess of unbound BG.

    Produced:

    Article Title: A Single Mechanism of Biogenesis, Initiated and Directed by PIWI Proteins, Explains piRNA Production in Most Animals
    Article Snippet: SNAP-tagged Mus musculus PIWIL1 (SNAP-MIWI) and PIWIL2 (SNAP-MILI) were produced in HEK293T cells from a lentiviral-transduced transgene (lentivirus backbone was a gift from Greiner Lab). .. SNAP-MIWI was labeled with SNAP substrate SNAP-Surface 549 (NEB, S9112) and resolved by electrophoresis through a 4–20% gradient SDS- polyacrylamide gel (Bio-Rad Laboratories, 5671085).

    Concentration Assay:

    Article Title: Comprehensive and quantitative mapping of RNA–protein interactions across a transcribed eukaryotic genome
    Article Snippet: Subsequently, Vts1(442–523)-SNAP-His6 was fluorescently labeled using SNAP-Surface549 (NEB). .. Concentration of labeled protein was measured using A 280 absorbance and was corrected for dye absorbance.

    Article Title: Ribonucleoprotein purification and characterization using RNA Mango
    Article Snippet: .. Yeast whole-cell extracts (500 µL) were first fluorescently tagged with the SNAP-Surface 549 tag (NEB) by incubating the extracts with 1 U/µL Murine RNase inhibitor (NEB), Protease Inhibitors (cOmplete mini at the recommended concentration, EDTA Free, Sigma), 5 mM DTT together with 5 µM SNAP-Surface 549 tag in Buffer B at RT for 30 min. .. Yeast whole-cell extracts (500 µL of extract either with or without SNAP label) were added to 200 µL of TO1-Dtb saturated agarose beads and incubated at 4°C for 1 h in a rotator.

    Article Title: A Single Mechanism of Biogenesis, Initiated and Directed by PIWI Proteins, Explains piRNA Production in Most Animals
    Article Snippet: SNAP-MIWI was labeled with SNAP substrate SNAP-Surface 549 (NEB, S9112) and resolved by electrophoresis through a 4–20% gradient SDS- polyacrylamide gel (Bio-Rad Laboratories, 5671085). .. The concentration of full-length SNAP-MIWI was determined by comparison to a standard curve of purified, SNAP- Surface 549 labeled SNAP protein (Typhoon FLA 7000; GE Lifesciences).

    Article Title: Synergistic assembly of human pre-spliceosomes across introns and exons
    Article Snippet: .. After centrifuging the lysate for 20 s at 12,000 × g , the nuclear pellet was resuspended in 450 µl Buffer C [20 mM Tris, 25% (v/v) glycerol, 0.42 M NaCl, 1.5 mM MgCl2 , 0.2 mM EDTA and 0.5 mM DTT, pH 7.9 at 4°C, supplemented with complete protease inhibitor cocktail (Roche, 04693159001)] and rapidly stirred in a 2 ml round-bottom microcentrifuge tube with a 12.7 × 3 mm stir bar for 30 min. After clarifying the lysate by centrifuging for 10 min at 12,000 × g , the SNAP-Surface 549 dye-benzylguanine conjugate (New England BioLabs, S9112S) was added to the supernatant at a final concentration of 200 nM and incubated for 30 min at 30°C. .. After labeling, the supernatant was dialyzed 2 times for 2 hr each against Buffer E (20 mM Tris, 20% (v/v) glycerol, 0.1 M KCl, 0.2 mM EDTA and 0.5 mM DTT, pH 7.9 at 4°C).

    Lysis:

    Article Title: A Single Mechanism of Biogenesis, Initiated and Directed by PIWI Proteins, Explains piRNA Production in Most Animals
    Article Snippet: Cells, washed twice with PBS, were homogenized in lysis buffer (30 mM HEPES-KOH pH 7.5, 100 mM potassium acetate, 3.5 mM magnesium acetate, 1 mM dithiothreitol, 20% (w/v) glycerol, 0.1% (v/v) Triton X-100, 1 mM 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride, 0.3 µM Aprotinin, 40 µM Bestatin, 10 µM E-64, 10 µM Leupeptin). .. SNAP-MIWI was labeled with SNAP substrate SNAP-Surface 549 (NEB, S9112) and resolved by electrophoresis through a 4–20% gradient SDS- polyacrylamide gel (Bio-Rad Laboratories, 5671085).

    Cross-linking Immunoprecipitation:

    Article Title: PD-L1:CD80 Cis-Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways
    Article Snippet: For , SNAP-tagged full length CD80 (SNAP–CD80), CD80 mutant (SNAP–CD80 I92R) or CD86 (SNAP–CD86) was transfected to either HEK293T cells or CLIP–PD-L1 transduced HEK293T cells using polyethylenimine. .. 24 h post transfection, cell was stained with a 1:1 mixture of SS549 (energy donor, NEB S9112S) and SSAF647 (energy acceptor) at 37°C, 5% CO2 for 30 min, and washed 3 times with 1x PBS (pH 7.4).

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    New England Biolabs bg549 surface
    Bg549 Surface, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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