ez link amine peg3 biotin  (Thermo Fisher)


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    Name:
    EZ Link Amine PEG3 Biotin
    Description:
    Thermo Scientific EZ Link Amine PEG3 Biotin is a water soluble biotin compounds containing a polyethylene glycol PEG spacer arm and a terminal primary amine for conjugation via EDC and other crosslinker methods Features of EZ Link Amine PEG3 Biotin • Biotinylation label molecules and surfaces for assays or affinity purification methods involving avidin or streptavidin probes and resins • Amine activated primary amine can be crosslinked to proteins and material surfaces using EDC and other crosslinkers • Pegylated polyethylene glycol PEG groups in spacer arm enhances water solubility of biotinylated molecules • Medium length spacer arm total length added to target is 22 9 angstroms Amine PEG2 Biotin and Amine PEG3 Biotin are the shorter two of three amine modified biotin compounds that contain polyethylene glycol PEG spacer arms The short PEG segments are hydrophilic and confer greater solubility to labeled proteins compared to reagents having only hydrocarbon spacers The primary amines of these pegylated biotin reagents can be conjugated to carboxyl groups on carboxy termini aspartate residues or glutamate residues using EDC Part No 22980 a water soluble carbodiimide crosslinker EDC activates carboxyl groups to bind to the NH2 group of the amino biotin forming an amide bond We manufacture biotin reagents to ensure the highest possible overall product integrity consistency and performance for the intended research applications Amino biotin compounds can be conjugated to functional groups of proteins and other molecules in a variety of ways The most common method is to crosslink the terminal primary amine to carboxyl groups using Carboxyl groups COOH occur in aspartate or glutamate residues and the carboxy terminus of polypeptides When activated with EDC Part No 22980 carboxylates react with amino NH2 groups to form amide bonds Carboxylate molecules and surface materials can be pre activated using EDC with Sulfo NHS Part No 24510 for subsequent reaction to primary amines see NHS ester Chemistry Related Products EZ Link Amine PEG2 Biotin
    Catalog Number:
    21347
    Price:
    None
    Applications:
    Protein Biology|Protein Labeling|Protein Labeling & Crosslinking
    Category:
    Labeling Detection Products
    Buy from Supplier


    Structured Review

    Thermo Fisher ez link amine peg3 biotin
    Schematic overview for the protein attached chromatin capture (PAtCh-Cap) method. Streptavidin coated magnetic beads are first conjugated with <t>amine-PEG3-biotin</t> (where PEG is polyethylene glycol). After standard cross-linking, chromatin isolation and DNA shearing procedures, 10% of the sample volume is removed and incubated with the pre-conjugated beads in the presence of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride). EDC reacts with protein carboxylate groups, forming unstable o -acylisourea activated esters that can undergo nucleophilic attack by the primary amines on the amine-PEG3-biotin prosthetics. This forms a covalent amide linkage with proteins in the chromatin complexes and releases an isourea by-product. Once bead-bound, these chromatin complexes can be subjected to additional chemical processing steps in parallel with bead-bound chromatin that was separately immunoprecipitated for the target of interest.
    Thermo Scientific EZ Link Amine PEG3 Biotin is a water soluble biotin compounds containing a polyethylene glycol PEG spacer arm and a terminal primary amine for conjugation via EDC and other crosslinker methods Features of EZ Link Amine PEG3 Biotin • Biotinylation label molecules and surfaces for assays or affinity purification methods involving avidin or streptavidin probes and resins • Amine activated primary amine can be crosslinked to proteins and material surfaces using EDC and other crosslinkers • Pegylated polyethylene glycol PEG groups in spacer arm enhances water solubility of biotinylated molecules • Medium length spacer arm total length added to target is 22 9 angstroms Amine PEG2 Biotin and Amine PEG3 Biotin are the shorter two of three amine modified biotin compounds that contain polyethylene glycol PEG spacer arms The short PEG segments are hydrophilic and confer greater solubility to labeled proteins compared to reagents having only hydrocarbon spacers The primary amines of these pegylated biotin reagents can be conjugated to carboxyl groups on carboxy termini aspartate residues or glutamate residues using EDC Part No 22980 a water soluble carbodiimide crosslinker EDC activates carboxyl groups to bind to the NH2 group of the amino biotin forming an amide bond We manufacture biotin reagents to ensure the highest possible overall product integrity consistency and performance for the intended research applications Amino biotin compounds can be conjugated to functional groups of proteins and other molecules in a variety of ways The most common method is to crosslink the terminal primary amine to carboxyl groups using Carboxyl groups COOH occur in aspartate or glutamate residues and the carboxy terminus of polypeptides When activated with EDC Part No 22980 carboxylates react with amino NH2 groups to form amide bonds Carboxylate molecules and surface materials can be pre activated using EDC with Sulfo NHS Part No 24510 for subsequent reaction to primary amines see NHS ester Chemistry Related Products EZ Link Amine PEG2 Biotin
    https://www.bioz.com/result/ez link amine peg3 biotin/product/Thermo Fisher
    Average 90 stars, based on 1 article reviews
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    ez link amine peg3 biotin - by Bioz Stars, 2020-01
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    Images

    1) Product Images from "PAtCh-Cap: input strategy for improving analysis of ChIP-exo data sets and beyond"

    Article Title: PAtCh-Cap: input strategy for improving analysis of ChIP-exo data sets and beyond

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkw741

    Schematic overview for the protein attached chromatin capture (PAtCh-Cap) method. Streptavidin coated magnetic beads are first conjugated with amine-PEG3-biotin (where PEG is polyethylene glycol). After standard cross-linking, chromatin isolation and DNA shearing procedures, 10% of the sample volume is removed and incubated with the pre-conjugated beads in the presence of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride). EDC reacts with protein carboxylate groups, forming unstable o -acylisourea activated esters that can undergo nucleophilic attack by the primary amines on the amine-PEG3-biotin prosthetics. This forms a covalent amide linkage with proteins in the chromatin complexes and releases an isourea by-product. Once bead-bound, these chromatin complexes can be subjected to additional chemical processing steps in parallel with bead-bound chromatin that was separately immunoprecipitated for the target of interest.
    Figure Legend Snippet: Schematic overview for the protein attached chromatin capture (PAtCh-Cap) method. Streptavidin coated magnetic beads are first conjugated with amine-PEG3-biotin (where PEG is polyethylene glycol). After standard cross-linking, chromatin isolation and DNA shearing procedures, 10% of the sample volume is removed and incubated with the pre-conjugated beads in the presence of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride). EDC reacts with protein carboxylate groups, forming unstable o -acylisourea activated esters that can undergo nucleophilic attack by the primary amines on the amine-PEG3-biotin prosthetics. This forms a covalent amide linkage with proteins in the chromatin complexes and releases an isourea by-product. Once bead-bound, these chromatin complexes can be subjected to additional chemical processing steps in parallel with bead-bound chromatin that was separately immunoprecipitated for the target of interest.

    Techniques Used: Magnetic Beads, Isolation, Incubation, Immunoprecipitation

    Related Articles

    Chromogenic Assay:

    Article Title: Coating of Biomaterial Scaffolds with the Collagen-Mimetic Peptide GFOGER for Bone Defect Repair
    Article Snippet: Samples contained 10-fold lower levels of endotoxin than the United States Food and Drug Administration’s recommended 0.5 EU/mL, as determined by the LAL chromogenic assay (Cambrex). .. For in vitro detection of GFOGER on PCL scaffolds, the carboxyl end of the GFOGER peptide was biotinylated using an EZ-Link® Amine-PEG3 -Biotin kit (Pierce Biotechnology), according to manufacturer’s instructions.

    Synthesized:

    Article Title: Engineering an antibody with picomolar affinity to DOTA chelates of multiple radionuclides for pretargeted radioimmunotherapy and imaging
    Article Snippet: .. DOTA-Bn-biotin ( ) was synthesized by dissolving Amine-PEG3 -Biotin purchased from Thermo Scientific (Rockford, IL) and S-2-(4-Isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-tetraacetic acid (p-SCN-Bn-DOTA) purchased from Macrocyclics in dimethyl sulfoxide (DMSO) with a 10 fold molar excess of triethylamine. .. The reaction mixture was vortexed at room temperature for 3 h, and then purified by high performance liquid chromatography (HPLC).

    Article Title: A modular IgG-scFv bispecific antibody topology
    Article Snippet: .. DOTA-biotin was synthesized by adding p-SCN-B n -DOTA ( S -2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-tetraacetic acid) purchased from Macrocyclics (Dallas, TX) to Amine-PEG3 -Biotin (Thermo Fisher Scientific) in dimethyl sulfoxide (DMSO) purchased from Sigma Aldrich (St. Louis, MO) with a 10-fold molar excess of triethylamine (TEA) purchased from VWR (West Chester, PA). .. The reaction mixture was vortexed at room temperature for 3 h, and purified by high-performance liquid chromatography (HPLC) on a C-18 reverse-phase column (Agilent Model 1100 HPLC, 1 × 25 cm, buffer A = 0.05% trifluoroacetic acid (TFA), buffer B = 0.0425% TFA in 80% acetonitrile, 2–100% B gradient for 98 min).

    Article Title: Site-Specific Incorporation of Quadricyclane into a Protein and Photocleavage of the Quadricyclane Ligation Adduct
    Article Snippet: .. Dithiocarbonate S1 ( ) was synthesized according to the route developed by Sletten et al. EZ-link amine-PEG3 -biotin from Thermo Scientific (53 mg) was transferred to a round bottom flask as a solution in water, concentrated to a solid on a rotary evaporator, and azeotroped from toluene (2 × 2 mL). ..

    Incubation:

    Article Title: Nanofiber Orientation and Surface Functionalization Modulate Human Mesenchymal Stem Cell Behavior In Vitro
    Article Snippet: Biotin was conjugated to the carboxyl end of the peptide using the EZ-Link® Amine-PEG3 -Biotin kit (Pierce Biotechnology), and unreacted biotin was removed by dialysis. .. To visualize the presence of the peptide on the nanofiber surface, the GFOGER-coated nanofiber mesh samples were incubated with 10 μg/mL of fluorescein-conjugated NeutrAvidin® (Molecular Probes) for 30 min at room temperature in the dark.

    Article Title: Graphene quantum dots prevent α-synucleinopathy in Parkinson’s disease
    Article Snippet: After 1 h of reaction with vigorous stirring, 25 mg of EZ-Link amine-PEG3 -biotin (cat. no. 21347; Thermo Scientific) was added to EDC-activated GQDs. .. For the binding assay between GQDs and α-syn fibrils, 5 mg ml−1 of α-syn fibrils were incubated with 5 mg ml−1 of biotinylated GQDs and streptavidin-conjugated 0.8 nm ultra-small gold particles (cat. no. 800.099, Aurion) for 1 h. Next, the streptavidin-conjugated ultra-small gold particles bound with high affinity to biotinylated-GQDs were enhanced with GoldEnhance EM Plus solution (cat.no.

    Article Title: PAtCh-Cap: input strategy for improving analysis of ChIP-exo data sets and beyond
    Article Snippet: Protein attached chromatin capture (PAtCh-Cap) for ChIP-exo Following two series of washes with 0.01 M PBS (pH 7.4), 50 μg of M-280 streptavidin coated Dynabeads (Thermo Fisher Scientific; equivalent to the number of beads utilized per reaction in the Active Motif ChIP-exo Kit) were conjugated with 10 μl of 50 nM EZ-link amine-PEG3-Biotin (Thermo Fisher Scientific; PEG = polyethylene glycol) in 0.01 M PBS (pH 7.4) at room temperature for 20 min. .. For each of the two replicates, the input sample (obtained as discussed above) was combined with 300 μl of 0.1 M EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) along with the pre-functionalized biotinylated magnetics beads and incubated in 10 ml 0.1 mM MES buffer (pH 5.0) for 3 h at room temperature on a mechanical rotator.

    Mass Spectrometry:

    Article Title: A modular IgG-scFv bispecific antibody topology
    Article Snippet: DOTA-biotin was synthesized by adding p-SCN-B n -DOTA ( S -2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-tetraacetic acid) purchased from Macrocyclics (Dallas, TX) to Amine-PEG3 -Biotin (Thermo Fisher Scientific) in dimethyl sulfoxide (DMSO) purchased from Sigma Aldrich (St. Louis, MO) with a 10-fold molar excess of triethylamine (TEA) purchased from VWR (West Chester, PA). .. Fractions containing DOTA-biotin were confirmed using matrix-assisted laser desorption/ionization–time of flight (MALDI-TOF) mass spectrometry (Applied Biosystems Model Voyager DE-STR).

    BIA-KA:

    Article Title: Coating of Biomaterial Scaffolds with the Collagen-Mimetic Peptide GFOGER for Bone Defect Repair
    Article Snippet: For in vitro detection of GFOGER on PCL scaffolds, the carboxyl end of the GFOGER peptide was biotinylated using an EZ-Link® Amine-PEG3 -Biotin kit (Pierce Biotechnology), according to manufacturer’s instructions. .. After dialysis, protein concentration was measured using a BCA Protein Assay kit (Pierce Biotechnology).

    Modification:

    Article Title: Comparison of the Interactions of Different Growth Factors and Glycosaminoglycans
    Article Snippet: .. Preparation of Heparin Biochip The preparation of biotinylated heparin was followed our previous protocol with minor modification [ ]: in 200 µL of H2 O, 2 mg of heparin and 2 mg of amine–PEG3–Biotin (Thermo Scientific, Waltham, MA, USA) were mixed with 10 mg of NaCNBH3 . .. The initial reaction was carried out at 70 °C for 24 h, and then a further 10 mg of NaCNBH3 was added to continue running the reaction for another 24 h. After completing the reaction, the mixture was desalted with a spin column (3000 molecular weight cut-off).

    High Performance Liquid Chromatography:

    Article Title: Engineering an antibody with picomolar affinity to DOTA chelates of multiple radionuclides for pretargeted radioimmunotherapy and imaging
    Article Snippet: DOTA-Bn-biotin ( ) was synthesized by dissolving Amine-PEG3 -Biotin purchased from Thermo Scientific (Rockford, IL) and S-2-(4-Isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-tetraacetic acid (p-SCN-Bn-DOTA) purchased from Macrocyclics in dimethyl sulfoxide (DMSO) with a 10 fold molar excess of triethylamine. .. The reaction mixture was vortexed at room temperature for 3 h, and then purified by high performance liquid chromatography (HPLC).

    Article Title: A modular IgG-scFv bispecific antibody topology
    Article Snippet: DOTA-biotin was synthesized by adding p-SCN-B n -DOTA ( S -2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-tetraacetic acid) purchased from Macrocyclics (Dallas, TX) to Amine-PEG3 -Biotin (Thermo Fisher Scientific) in dimethyl sulfoxide (DMSO) purchased from Sigma Aldrich (St. Louis, MO) with a 10-fold molar excess of triethylamine (TEA) purchased from VWR (West Chester, PA). .. The reaction mixture was vortexed at room temperature for 3 h, and purified by high-performance liquid chromatography (HPLC) on a C-18 reverse-phase column (Agilent Model 1100 HPLC, 1 × 25 cm, buffer A = 0.05% trifluoroacetic acid (TFA), buffer B = 0.0425% TFA in 80% acetonitrile, 2–100% B gradient for 98 min).

    Conjugation Assay:

    Article Title: Graphene quantum dots prevent α-synucleinopathy in Parkinson’s disease
    Article Snippet: GQDs (50 mg) were dissolved in conjugation buffer (pH 4.7), and 12.5 mg of EDC reagent ( N -(3-dimethylaminopropyl)- N ′′-ethylcarbodiimide hydrochloride; cat. no. 03449, Sigma-Aldrich) was subsequently added to replace the carboxyl groups. .. After 1 h of reaction with vigorous stirring, 25 mg of EZ-Link amine-PEG3 -biotin (cat. no. 21347; Thermo Scientific) was added to EDC-activated GQDs.

    Flow Cytometry:

    Article Title: Engineering an antibody with picomolar affinity to DOTA chelates of multiple radionuclides for pretargeted radioimmunotherapy and imaging
    Article Snippet: DOTA-Bn-biotin ( ) was synthesized by dissolving Amine-PEG3 -Biotin purchased from Thermo Scientific (Rockford, IL) and S-2-(4-Isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-tetraacetic acid (p-SCN-Bn-DOTA) purchased from Macrocyclics in dimethyl sulfoxide (DMSO) with a 10 fold molar excess of triethylamine. .. Flow through was monitored by absorbance detection at 280 nanometers.

    Article Title: A modular IgG-scFv bispecific antibody topology
    Article Snippet: DOTA-biotin was synthesized by adding p-SCN-B n -DOTA ( S -2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-tetraacetic acid) purchased from Macrocyclics (Dallas, TX) to Amine-PEG3 -Biotin (Thermo Fisher Scientific) in dimethyl sulfoxide (DMSO) purchased from Sigma Aldrich (St. Louis, MO) with a 10-fold molar excess of triethylamine (TEA) purchased from VWR (West Chester, PA). .. Flow through was monitored by absorbance detection at 280 nm.

    Article Title: Major Differences between the Self-Assembly and Seeding Behavior of Heparin-Induced and in Vitro Phosphorylated Tau and Their Modulation by Potential Inhibitors
    Article Snippet: Biotinylated heparin was prepared by reaction with amine-PEG3 -Biotin (Pierce, Rockford, IL) in the presence of NaCNBH3 as described previously. .. Briefly, 20 μL of 0.1 mg mL–1 of the biotinylated heparin in HBS-EP running buffer (0.01 M HEPES, 0.15 M NaCl, 3 mM EDTA, and 0.005% surfactant P20, pH 7.4) were injected over flow cells 2, 3, and 4 of the SA chip at a flow rate of 10 μL/min.

    Infection:

    Article Title: BmGPAC, an Antigen Capture Assay for Detection of Active Babesia microti Infection
    Article Snippet: This antiserum recognizes a single protein in Babesia -infected RBCs with the molecular weight expected for BmGPI12. .. Carboxyl groups of the IgG molecules were biotinylated with an EZ-Link amine-PEG3-biotin kit (Thermo Fisher Scientific), according to the manufacturer's instructions.

    Protein Concentration:

    Article Title: Coating of Biomaterial Scaffolds with the Collagen-Mimetic Peptide GFOGER for Bone Defect Repair
    Article Snippet: For in vitro detection of GFOGER on PCL scaffolds, the carboxyl end of the GFOGER peptide was biotinylated using an EZ-Link® Amine-PEG3 -Biotin kit (Pierce Biotechnology), according to manufacturer’s instructions. .. After dialysis, protein concentration was measured using a BCA Protein Assay kit (Pierce Biotechnology).

    Injection:

    Article Title: Major Differences between the Self-Assembly and Seeding Behavior of Heparin-Induced and in Vitro Phosphorylated Tau and Their Modulation by Potential Inhibitors
    Article Snippet: Biotinylated heparin was prepared by reaction with amine-PEG3 -Biotin (Pierce, Rockford, IL) in the presence of NaCNBH3 as described previously. .. Briefly, 20 μL of 0.1 mg mL–1 of the biotinylated heparin in HBS-EP running buffer (0.01 M HEPES, 0.15 M NaCl, 3 mM EDTA, and 0.005% surfactant P20, pH 7.4) were injected over flow cells 2, 3, and 4 of the SA chip at a flow rate of 10 μL/min.

    Binding Assay:

    Article Title: Binding Properties of the Transforming Growth Factor-β Coreceptor Betaglycan: Proposed Mechanism for Potentiation of Receptor Complex Assembly and Signaling
    Article Snippet: Paragraph title: SPR Binding Measurements ... All SPR experiments, except those reported in Table 1 of the Supporting Information , were performed using TGF-βs biotinylated in 25 mM MES (pH 4.8) with a 100-fold molar excess of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide·HCl (EDC), a 25-fold molar excess of N -hydroxysulfosuccinimide (Sulfo-NHS), and a 100-fold molar excess of EZ-Link Amine-PEG3 -biotin (Pierce, Rockford, IL).

    Article Title: Graphene quantum dots prevent α-synucleinopathy in Parkinson’s disease
    Article Snippet: Paragraph title: Biotinylation of GQDs and binding assay. ... After 1 h of reaction with vigorous stirring, 25 mg of EZ-Link amine-PEG3 -biotin (cat. no. 21347; Thermo Scientific) was added to EDC-activated GQDs.

    Molecular Weight:

    Article Title: BmGPAC, an Antigen Capture Assay for Detection of Active Babesia microti Infection
    Article Snippet: This antiserum recognizes a single protein in Babesia -infected RBCs with the molecular weight expected for BmGPI12. .. Carboxyl groups of the IgG molecules were biotinylated with an EZ-Link amine-PEG3-biotin kit (Thermo Fisher Scientific), according to the manufacturer's instructions.

    Article Title: Comparison of the Interactions of Different Growth Factors and Glycosaminoglycans
    Article Snippet: Preparation of Heparin Biochip The preparation of biotinylated heparin was followed our previous protocol with minor modification [ ]: in 200 µL of H2 O, 2 mg of heparin and 2 mg of amine–PEG3–Biotin (Thermo Scientific, Waltham, MA, USA) were mixed with 10 mg of NaCNBH3 . .. The initial reaction was carried out at 70 °C for 24 h, and then a further 10 mg of NaCNBH3 was added to continue running the reaction for another 24 h. After completing the reaction, the mixture was desalted with a spin column (3000 molecular weight cut-off).

    Article Title: Copper regulates the interactions of antimicrobial piscidin peptides from fish mast cells with formyl peptide receptors and heparin
    Article Snippet: Heparin (2 mg) and amine–PEG3–Biotin (2 mg, Pierce) were dissolved in 200 μl of H2 O and mixed with 10 mg of NaCNBH3 . .. The reaction mixture was heated at 70 °C for 24 h; after that, a further 10 mg of NaCNBH3 was added, and the reaction was carried for another 24 h. After cooling to room temperature, the mixture was desalted with a spin column (3000 molecular weight cutoff).

    Article Title: Coating of Biomaterial Scaffolds with the Collagen-Mimetic Peptide GFOGER for Bone Defect Repair
    Article Snippet: For in vitro detection of GFOGER on PCL scaffolds, the carboxyl end of the GFOGER peptide was biotinylated using an EZ-Link® Amine-PEG3 -Biotin kit (Pierce Biotechnology), according to manufacturer’s instructions. .. Unreacted biotin was removed via overnight dialysis against PBS using a Slide-A-Lyzer Dialysis Cassette with a molecular weight cut-off of 3500 (Thermo Scientific).

    Article Title: Graphene quantum dots prevent α-synucleinopathy in Parkinson’s disease
    Article Snippet: After 1 h of reaction with vigorous stirring, 25 mg of EZ-Link amine-PEG3 -biotin (cat. no. 21347; Thermo Scientific) was added to EDC-activated GQDs. .. 2114, Nanoprobes) for 5 min. Non-reacted solution was removed by a 100 kDa molecular weight cut-off spin column (cat. no. UFC510024, Millipore-Sigma), followed by TEM analysis.

    Ion Exchange Chromatography:

    Article Title: Binding Properties of the Transforming Growth Factor-β Coreceptor Betaglycan: Proposed Mechanism for Potentiation of Receptor Complex Assembly and Signaling
    Article Snippet: All SPR experiments, except those reported in Table 1 of the Supporting Information , were performed using TGF-βs biotinylated in 25 mM MES (pH 4.8) with a 100-fold molar excess of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide·HCl (EDC), a 25-fold molar excess of N -hydroxysulfosuccinimide (Sulfo-NHS), and a 100-fold molar excess of EZ-Link Amine-PEG3 -biotin (Pierce, Rockford, IL). .. The biotinylation reactions were quenched with 10 volumes of 100 mM acetic acid, and the biotinylated TGF-βs were isolated by ion exchange chromatography (Source S, GE Healthcare) at pH 4.0 in 25 mM NaOAc and 30% isopropanol.

    Fluorescence:

    Article Title: Nanofiber Orientation and Surface Functionalization Modulate Human Mesenchymal Stem Cell Behavior In Vitro
    Article Snippet: Biotin was conjugated to the carboxyl end of the peptide using the EZ-Link® Amine-PEG3 -Biotin kit (Pierce Biotechnology), and unreacted biotin was removed by dialysis. .. After rinsing with excess PBS, images were taken on an inverted fluorescence microscope (Axio Observer.Z1; Carl Zeiss) using a fluorescein isothiocyanate filter.

    Magnetic Beads:

    Article Title: PAtCh-Cap: input strategy for improving analysis of ChIP-exo data sets and beyond
    Article Snippet: Protein attached chromatin capture (PAtCh-Cap) for ChIP-exo Following two series of washes with 0.01 M PBS (pH 7.4), 50 μg of M-280 streptavidin coated Dynabeads (Thermo Fisher Scientific; equivalent to the number of beads utilized per reaction in the Active Motif ChIP-exo Kit) were conjugated with 10 μl of 50 nM EZ-link amine-PEG3-Biotin (Thermo Fisher Scientific; PEG = polyethylene glycol) in 0.01 M PBS (pH 7.4) at room temperature for 20 min. .. These beads were selected as they have the same size and core material composition as the protein G coated magnetic beads utilized in the Active Motif ChIP-exo Kit.

    Isolation:

    Article Title: Binding Properties of the Transforming Growth Factor-β Coreceptor Betaglycan: Proposed Mechanism for Potentiation of Receptor Complex Assembly and Signaling
    Article Snippet: All SPR experiments, except those reported in Table 1 of the Supporting Information , were performed using TGF-βs biotinylated in 25 mM MES (pH 4.8) with a 100-fold molar excess of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide·HCl (EDC), a 25-fold molar excess of N -hydroxysulfosuccinimide (Sulfo-NHS), and a 100-fold molar excess of EZ-Link Amine-PEG3 -biotin (Pierce, Rockford, IL). .. The biotinylation reactions were quenched with 10 volumes of 100 mM acetic acid, and the biotinylated TGF-βs were isolated by ion exchange chromatography (Source S, GE Healthcare) at pH 4.0 in 25 mM NaOAc and 30% isopropanol.

    Microscopy:

    Article Title: Nanofiber Orientation and Surface Functionalization Modulate Human Mesenchymal Stem Cell Behavior In Vitro
    Article Snippet: Biotin was conjugated to the carboxyl end of the peptide using the EZ-Link® Amine-PEG3 -Biotin kit (Pierce Biotechnology), and unreacted biotin was removed by dialysis. .. After rinsing with excess PBS, images were taken on an inverted fluorescence microscope (Axio Observer.Z1; Carl Zeiss) using a fluorescein isothiocyanate filter.

    Purification:

    Article Title: Engineering an antibody with picomolar affinity to DOTA chelates of multiple radionuclides for pretargeted radioimmunotherapy and imaging
    Article Snippet: DOTA-Bn-biotin ( ) was synthesized by dissolving Amine-PEG3 -Biotin purchased from Thermo Scientific (Rockford, IL) and S-2-(4-Isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-tetraacetic acid (p-SCN-Bn-DOTA) purchased from Macrocyclics in dimethyl sulfoxide (DMSO) with a 10 fold molar excess of triethylamine. .. The reaction mixture was vortexed at room temperature for 3 h, and then purified by high performance liquid chromatography (HPLC).

    Article Title: Changes in Metabolic Chemical Reporter Structure Yield a Selective Probe of O-GlcNAc Modification
    Article Snippet: EZ-Link Amine PEG3 -Biotin (0.460 mg, 0.111 mmol) (Thermo Scientific) was added and reaction let stir for 18 h upon which time solvent was removed by vacuum. .. The resulting crude mixture was purified by RP-HPLC over a C18 semipreparative column (The Nest Group) using a 5.5–44% B linear gradient over 10 min before switching to a 44–100% B linear gradient over 40 min, t R = 18 min (buffer A: 0.1% TFA in water, buffer B: 0.1% TFA, 90% ACN in water) and lyophilized to afford the pure product as an orange solid (0.022 g, 31%): MALDI-MS calculated for C34 H40 N6 O8 S (oxidized at the biotin cysteine) [M + Na]+ 719.2839, found 719.2656.

    Article Title: BmGPAC, an Antigen Capture Assay for Detection of Active Babesia microti Infection
    Article Snippet: IgG was purified from the antiserum by affinity chromatography on a HiTrap protein G column (GE Healthcare), as recommended by the manufacturer. .. Carboxyl groups of the IgG molecules were biotinylated with an EZ-Link amine-PEG3-biotin kit (Thermo Fisher Scientific), according to the manufacturer's instructions.

    Article Title: A modular IgG-scFv bispecific antibody topology
    Article Snippet: DOTA-biotin was synthesized by adding p-SCN-B n -DOTA ( S -2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-tetraacetic acid) purchased from Macrocyclics (Dallas, TX) to Amine-PEG3 -Biotin (Thermo Fisher Scientific) in dimethyl sulfoxide (DMSO) purchased from Sigma Aldrich (St. Louis, MO) with a 10-fold molar excess of triethylamine (TEA) purchased from VWR (West Chester, PA). .. The reaction mixture was vortexed at room temperature for 3 h, and purified by high-performance liquid chromatography (HPLC) on a C-18 reverse-phase column (Agilent Model 1100 HPLC, 1 × 25 cm, buffer A = 0.05% trifluoroacetic acid (TFA), buffer B = 0.0425% TFA in 80% acetonitrile, 2–100% B gradient for 98 min).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Graphene quantum dots prevent α-synucleinopathy in Parkinson’s disease
    Article Snippet: After 1 h of reaction with vigorous stirring, 25 mg of EZ-Link amine-PEG3 -biotin (cat. no. 21347; Thermo Scientific) was added to EDC-activated GQDs. .. For the binding assay between GQDs and α-syn fibrils, 5 mg ml−1 of α-syn fibrils were incubated with 5 mg ml−1 of biotinylated GQDs and streptavidin-conjugated 0.8 nm ultra-small gold particles (cat. no. 800.099, Aurion) for 1 h. Next, the streptavidin-conjugated ultra-small gold particles bound with high affinity to biotinylated-GQDs were enhanced with GoldEnhance EM Plus solution (cat.no.

    Chromatin Immunoprecipitation:

    Article Title: Binding Properties of the Transforming Growth Factor-β Coreceptor Betaglycan: Proposed Mechanism for Potentiation of Receptor Complex Assembly and Signaling
    Article Snippet: All SPR experiments, except those reported in Table 1 of the Supporting Information , were performed using TGF-βs biotinylated in 25 mM MES (pH 4.8) with a 100-fold molar excess of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide·HCl (EDC), a 25-fold molar excess of N -hydroxysulfosuccinimide (Sulfo-NHS), and a 100-fold molar excess of EZ-Link Amine-PEG3 -biotin (Pierce, Rockford, IL). .. Streptavidin was coupled to a CM5 sensor chip (GE Healthcare) by activation with EDC/NHS to 3000–5000 resonance units (RUs).

    Article Title: Comparison of the Interactions of Different Growth Factors and Glycosaminoglycans
    Article Snippet: Preparation of Heparin Biochip The preparation of biotinylated heparin was followed our previous protocol with minor modification [ ]: in 200 µL of H2 O, 2 mg of heparin and 2 mg of amine–PEG3–Biotin (Thermo Scientific, Waltham, MA, USA) were mixed with 10 mg of NaCNBH3 . .. Biotinylated heparin was freeze-dried for chip preparation.

    Article Title: Major Differences between the Self-Assembly and Seeding Behavior of Heparin-Induced and in Vitro Phosphorylated Tau and Their Modulation by Potential Inhibitors
    Article Snippet: Biotinylated heparin was prepared by reaction with amine-PEG3 -Biotin (Pierce, Rockford, IL) in the presence of NaCNBH3 as described previously. .. The biotinylated heparin was immobilized on a streptavidin (SA) chip (GE Healthcare) based on the manufacturer’s protocol.

    Article Title: Copper regulates the interactions of antimicrobial piscidin peptides from fish mast cells with formyl peptide receptors and heparin
    Article Snippet: Heparin (2 mg) and amine–PEG3–Biotin (2 mg, Pierce) were dissolved in 200 μl of H2 O and mixed with 10 mg of NaCNBH3 . .. Biotinylated heparin was freeze-dried for chip preparation.

    Article Title: PAtCh-Cap: input strategy for improving analysis of ChIP-exo data sets and beyond
    Article Snippet: .. Protein attached chromatin capture (PAtCh-Cap) for ChIP-exo Following two series of washes with 0.01 M PBS (pH 7.4), 50 μg of M-280 streptavidin coated Dynabeads (Thermo Fisher Scientific; equivalent to the number of beads utilized per reaction in the Active Motif ChIP-exo Kit) were conjugated with 10 μl of 50 nM EZ-link amine-PEG3-Biotin (Thermo Fisher Scientific; PEG = polyethylene glycol) in 0.01 M PBS (pH 7.4) at room temperature for 20 min. .. These beads were selected as they have the same size and core material composition as the protein G coated magnetic beads utilized in the Active Motif ChIP-exo Kit.

    SPR Assay:

    Article Title: Binding Properties of the Transforming Growth Factor-β Coreceptor Betaglycan: Proposed Mechanism for Potentiation of Receptor Complex Assembly and Signaling
    Article Snippet: .. All SPR experiments, except those reported in Table 1 of the Supporting Information , were performed using TGF-βs biotinylated in 25 mM MES (pH 4.8) with a 100-fold molar excess of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide·HCl (EDC), a 25-fold molar excess of N -hydroxysulfosuccinimide (Sulfo-NHS), and a 100-fold molar excess of EZ-Link Amine-PEG3 -biotin (Pierce, Rockford, IL). .. SPR experiments, reported in Table 1 of the Supporting Information , were performed using TGF-β2 biotinylated by prebinding it to BGO-ZP in 10 mM sodium phosphate and 140 mM NaCl (pH 7.5) followed by treatment with 1 molar equivalant of sulfo-NHS-LC-LC-Biotin (Pierce).

    Article Title: Major Differences between the Self-Assembly and Seeding Behavior of Heparin-Induced and in Vitro Phosphorylated Tau and Their Modulation by Potential Inhibitors
    Article Snippet: Paragraph title: Surface Plasmon Resonance ... Biotinylated heparin was prepared by reaction with amine-PEG3 -Biotin (Pierce, Rockford, IL) in the presence of NaCNBH3 as described previously.

    Affinity Chromatography:

    Article Title: BmGPAC, an Antigen Capture Assay for Detection of Active Babesia microti Infection
    Article Snippet: IgG was purified from the antiserum by affinity chromatography on a HiTrap protein G column (GE Healthcare), as recommended by the manufacturer. .. Carboxyl groups of the IgG molecules were biotinylated with an EZ-Link amine-PEG3-biotin kit (Thermo Fisher Scientific), according to the manufacturer's instructions.

    In Vitro:

    Article Title: Coating of Biomaterial Scaffolds with the Collagen-Mimetic Peptide GFOGER for Bone Defect Repair
    Article Snippet: .. For in vitro detection of GFOGER on PCL scaffolds, the carboxyl end of the GFOGER peptide was biotinylated using an EZ-Link® Amine-PEG3 -Biotin kit (Pierce Biotechnology), according to manufacturer’s instructions. .. Unreacted biotin was removed via overnight dialysis against PBS using a Slide-A-Lyzer Dialysis Cassette with a molecular weight cut-off of 3500 (Thermo Scientific).

    Evaporation:

    Article Title: Graphene quantum dots prevent α-synucleinopathy in Parkinson’s disease
    Article Snippet: After 1 h of reaction with vigorous stirring, 25 mg of EZ-Link amine-PEG3 -biotin (cat. no. 21347; Thermo Scientific) was added to EDC-activated GQDs. .. The solution was then subjected to dialysis and rotary evaporation to yield the final product in powder form.

    Activation Assay:

    Article Title: Binding Properties of the Transforming Growth Factor-β Coreceptor Betaglycan: Proposed Mechanism for Potentiation of Receptor Complex Assembly and Signaling
    Article Snippet: All SPR experiments, except those reported in Table 1 of the Supporting Information , were performed using TGF-βs biotinylated in 25 mM MES (pH 4.8) with a 100-fold molar excess of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide·HCl (EDC), a 25-fold molar excess of N -hydroxysulfosuccinimide (Sulfo-NHS), and a 100-fold molar excess of EZ-Link Amine-PEG3 -biotin (Pierce, Rockford, IL). .. Streptavidin was coupled to a CM5 sensor chip (GE Healthcare) by activation with EDC/NHS to 3000–5000 resonance units (RUs).

    Transmission Electron Microscopy:

    Article Title: Graphene quantum dots prevent α-synucleinopathy in Parkinson’s disease
    Article Snippet: After 1 h of reaction with vigorous stirring, 25 mg of EZ-Link amine-PEG3 -biotin (cat. no. 21347; Thermo Scientific) was added to EDC-activated GQDs. .. 2114, Nanoprobes) for 5 min. Non-reacted solution was removed by a 100 kDa molecular weight cut-off spin column (cat. no. UFC510024, Millipore-Sigma), followed by TEM analysis.

    Recombinant:

    Article Title: BmGPAC, an Antigen Capture Assay for Detection of Active Babesia microti Infection
    Article Snippet: The endpoint dilution titer of the resulting antiserum was greater than 108 when tested against recombinant BmGPI12. .. Carboxyl groups of the IgG molecules were biotinylated with an EZ-Link amine-PEG3-biotin kit (Thermo Fisher Scientific), according to the manufacturer's instructions.

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    Thermo Fisher ez link amine peg3 biotin
    Schematic overview for the protein attached chromatin capture (PAtCh-Cap) method. Streptavidin coated magnetic beads are first conjugated with <t>amine-PEG3-biotin</t> (where PEG is polyethylene glycol). After standard cross-linking, chromatin isolation and DNA shearing procedures, 10% of the sample volume is removed and incubated with the pre-conjugated beads in the presence of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride). EDC reacts with protein carboxylate groups, forming unstable o -acylisourea activated esters that can undergo nucleophilic attack by the primary amines on the amine-PEG3-biotin prosthetics. This forms a covalent amide linkage with proteins in the chromatin complexes and releases an isourea by-product. Once bead-bound, these chromatin complexes can be subjected to additional chemical processing steps in parallel with bead-bound chromatin that was separately immunoprecipitated for the target of interest.
    Ez Link Amine Peg3 Biotin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ez link amine peg3 biotin - by Bioz Stars, 2020-01
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    Schematic overview for the protein attached chromatin capture (PAtCh-Cap) method. Streptavidin coated magnetic beads are first conjugated with amine-PEG3-biotin (where PEG is polyethylene glycol). After standard cross-linking, chromatin isolation and DNA shearing procedures, 10% of the sample volume is removed and incubated with the pre-conjugated beads in the presence of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride). EDC reacts with protein carboxylate groups, forming unstable o -acylisourea activated esters that can undergo nucleophilic attack by the primary amines on the amine-PEG3-biotin prosthetics. This forms a covalent amide linkage with proteins in the chromatin complexes and releases an isourea by-product. Once bead-bound, these chromatin complexes can be subjected to additional chemical processing steps in parallel with bead-bound chromatin that was separately immunoprecipitated for the target of interest.

    Journal: Nucleic Acids Research

    Article Title: PAtCh-Cap: input strategy for improving analysis of ChIP-exo data sets and beyond

    doi: 10.1093/nar/gkw741

    Figure Lengend Snippet: Schematic overview for the protein attached chromatin capture (PAtCh-Cap) method. Streptavidin coated magnetic beads are first conjugated with amine-PEG3-biotin (where PEG is polyethylene glycol). After standard cross-linking, chromatin isolation and DNA shearing procedures, 10% of the sample volume is removed and incubated with the pre-conjugated beads in the presence of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride). EDC reacts with protein carboxylate groups, forming unstable o -acylisourea activated esters that can undergo nucleophilic attack by the primary amines on the amine-PEG3-biotin prosthetics. This forms a covalent amide linkage with proteins in the chromatin complexes and releases an isourea by-product. Once bead-bound, these chromatin complexes can be subjected to additional chemical processing steps in parallel with bead-bound chromatin that was separately immunoprecipitated for the target of interest.

    Article Snippet: Protein attached chromatin capture (PAtCh-Cap) for ChIP-exo Following two series of washes with 0.01 M PBS (pH 7.4), 50 μg of M-280 streptavidin coated Dynabeads (Thermo Fisher Scientific; equivalent to the number of beads utilized per reaction in the Active Motif ChIP-exo Kit) were conjugated with 10 μl of 50 nM EZ-link amine-PEG3-Biotin (Thermo Fisher Scientific; PEG = polyethylene glycol) in 0.01 M PBS (pH 7.4) at room temperature for 20 min.

    Techniques: Magnetic Beads, Isolation, Incubation, Immunoprecipitation