ez link phosphine peg3 biotin  (Thermo Fisher)


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    Name:
    EZ Link Phosphine PEG3 Biotin
    Description:
    Thermo Scientific Pierce EZ Link Phosphine PEG3 Biotin is a biotinylation reagent for labeling azide containing molecules which enables biotin based detection and affinity purification of molecules via Staudinger ligation strategies Features of EZ Link Phosphine PEG3 Biotin • Soluble easily dissolves in water miscible solvents e g DMSO for subsequent dilution in aqueous reaction mixtures with cell lysates and other biological samples • Compatible reaction chemistry occurs effectively in simple buffer conditions requires no accessory reagents such as copper or reducing agents • Chemoselective the phosphine reactive group is specific in biological samples for bioorthogonal azide tagged molecules ensuring that biotinylation is specific • PEG spacer polyethylene glycol spacer arm helps maintain solubility of labeled molecules and decreases steric hindrance for affinity binding to avidin streptavidin or NeutrAvidin Protein When used in combination with azide labeling strategies this compound enables detection or affinity purification of protein interactions and post translational modifications using streptavidin probes or streptavidin agarose resins The phosphine group of Phosphine PEG3 Biotin conjugates to azide groups by the Staudinger reaction mechanism Azide groups can be introduced into proteins or other cellular targets through in vivo labeling with azide tagged derivatives of naturally occurring metabolic building blocks Because neither phosphines nor azides are present in biological systems they comprise a chemoselective mutually specific ligation pair for labeling and conjugation
    Catalog Number:
    88901
    Price:
    None
    Applications:
    Protein Biology|Protein Labeling|Protein Labeling & Crosslinking
    Category:
    Labeling Detection Products
    Buy from Supplier


    Structured Review

    Thermo Fisher ez link phosphine peg3 biotin
    Thermo Scientific Pierce EZ Link Phosphine PEG3 Biotin is a biotinylation reagent for labeling azide containing molecules which enables biotin based detection and affinity purification of molecules via Staudinger ligation strategies Features of EZ Link Phosphine PEG3 Biotin • Soluble easily dissolves in water miscible solvents e g DMSO for subsequent dilution in aqueous reaction mixtures with cell lysates and other biological samples • Compatible reaction chemistry occurs effectively in simple buffer conditions requires no accessory reagents such as copper or reducing agents • Chemoselective the phosphine reactive group is specific in biological samples for bioorthogonal azide tagged molecules ensuring that biotinylation is specific • PEG spacer polyethylene glycol spacer arm helps maintain solubility of labeled molecules and decreases steric hindrance for affinity binding to avidin streptavidin or NeutrAvidin Protein When used in combination with azide labeling strategies this compound enables detection or affinity purification of protein interactions and post translational modifications using streptavidin probes or streptavidin agarose resins The phosphine group of Phosphine PEG3 Biotin conjugates to azide groups by the Staudinger reaction mechanism Azide groups can be introduced into proteins or other cellular targets through in vivo labeling with azide tagged derivatives of naturally occurring metabolic building blocks Because neither phosphines nor azides are present in biological systems they comprise a chemoselective mutually specific ligation pair for labeling and conjugation
    https://www.bioz.com/result/ez link phosphine peg3 biotin/product/Thermo Fisher
    Average 88 stars, based on 1 article reviews
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    ez link phosphine peg3 biotin - by Bioz Stars, 2020-01
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    Related Articles

    Nucleic Acid Electrophoresis:

    Article Title: Mechanism of oxidant-induced mistranslation by threonyl-tRNA synthetase
    Article Snippet: Biotinylation of DAz-2-labeled E. coli ThrRS and western blot analysis DAz-2-labeled ThrRS was conjugated to biotin via Staudinger ligation with 0.25 mM phosphine-PEG3-biotin (Thermo Scientific) at 37°C for 2 h. Biotinylation reactions were terminated by the addition of 1 ml of cold acetone and were kept in −80°C for 20 min. .. Protein pellet was washed once by 200 μl acetone then resuspended in 2x sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) loading dye.

    Article Title: Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis
    Article Snippet: The total protein of each lysate was quantified by BCA assay and the lysates were incubated with 500 nM of phosphine–PEG3 -biotin (Pierce, Rockford, IL, USA) for 6 h at 37 °C. .. The proteins from each sample were mixed with 1 x sodium dodecyl sulfate (SDS) gel-loading buffer and boiled for 5 min. 20 μg of proteins were separated by 10% SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membranes.

    Centrifugation:

    Article Title: Bioorthogonal labeling cell-surface proteins expressed in pancreatic cancer cells to identify potential diagnostic/therapeutic biomarkers
    Article Snippet: After 72 h, the cells were washed twice with phosphate-buffered saline (PBS) then treated with culture medium containing 200 µM of either EZ-Link Phosphine-PEG3 -Biotin (Thermo Fisher Scientific, 88901) or DBCO-S-S-PEG3-Biotin (Click Chemistry Tools, A112) conjugate suspended in DMSO to selectively tag glycoproteins on the surface of live cells. .. The cell suspensions from each sample (azido sugar or control sugar) were pooled, collected by centrifugation, washed twice with PBS, and lysed with RIPA buffer (1 mM EDTA, 1% NP-40, 0.5% deoxycholate, 0.1% SDS in PBS) followed by sonication.

    Article Title: Cell Labeling and Tracking Method without Distorted Signals by Phagocytosis of Macrophages
    Article Snippet: The insoluble debris was removed by centrifugation for 10 min at 3,000×g. .. Then, 20 µL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3 -biotin (2 µL, 5 mM in DPBS) (Pierce) for 6 h at 37 o C. Loading buffer was added to each sample, and samples were loaded onto 10% SPS-PAGE gel after heating at 95 o C. Proteins were transferred to Hybond P membrane (Amercham, St. Albans, UK), and the membrane was blocked with 5% bovine serum albumin in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH 7.4) for 2 h. The membrane was then incubated with streptavidin-horseradish peroxidase (diluted 1:2,000 in TBST) (Pierce) overnight at 4 o C. The membrane was rinsed three times with TBST and developed by using ECL Western Blotting Substrate (Pierce).

    Article Title: Physiological Effects of Ac4ManNAz and Optimization of Metabolic Labeling for Cell Tracking
    Article Snippet: The insoluble debris was removed by centrifugation for 10 min at 3,000×g. .. Then, 20 μL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3-biotin (2 μL, 5 mM in DPBS) (Pierce) for 6 h at 37 °C.

    Sonication:

    Article Title: Bioorthogonal labeling cell-surface proteins expressed in pancreatic cancer cells to identify potential diagnostic/therapeutic biomarkers
    Article Snippet: After 72 h, the cells were washed twice with phosphate-buffered saline (PBS) then treated with culture medium containing 200 µM of either EZ-Link Phosphine-PEG3 -Biotin (Thermo Fisher Scientific, 88901) or DBCO-S-S-PEG3-Biotin (Click Chemistry Tools, A112) conjugate suspended in DMSO to selectively tag glycoproteins on the surface of live cells. .. The cell suspensions from each sample (azido sugar or control sugar) were pooled, collected by centrifugation, washed twice with PBS, and lysed with RIPA buffer (1 mM EDTA, 1% NP-40, 0.5% deoxycholate, 0.1% SDS in PBS) followed by sonication.

    Article Title: Cell Labeling and Tracking Method without Distorted Signals by Phagocytosis of Macrophages
    Article Snippet: Sonicated lysates were incubated at 4 °C for 30 min to further solubilize proteins. .. Then, 20 µL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3 -biotin (2 µL, 5 mM in DPBS) (Pierce) for 6 h at 37 o C. Loading buffer was added to each sample, and samples were loaded onto 10% SPS-PAGE gel after heating at 95 o C. Proteins were transferred to Hybond P membrane (Amercham, St. Albans, UK), and the membrane was blocked with 5% bovine serum albumin in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH 7.4) for 2 h. The membrane was then incubated with streptavidin-horseradish peroxidase (diluted 1:2,000 in TBST) (Pierce) overnight at 4 o C. The membrane was rinsed three times with TBST and developed by using ECL Western Blotting Substrate (Pierce).

    Article Title: The Metabolic Chemical Reporter 6‑Azido-6-deoxy-glucose Further Reveals the Substrate Promiscuity of O‑GlcNAc Transferase and Catalyzes the Discovery of Intracellular Protein Modification by O‑Glucose
    Article Snippet: .. The mixture was sonicated in a bath sonicator to ensure complete dissolution, and then 73 μ L of H2 O was added, and 2 μ L of 10 mM EZ-Link Phosphine-PEG3 -Biotin (10 mM in DMSO, Thermo Scientific) was added and then incubated for 3 h at 37 °C and quenched with chloroform/ methanol precipitation. ..

    Ligation:

    Article Title: Mechanism of oxidant-induced mistranslation by threonyl-tRNA synthetase
    Article Snippet: .. Biotinylation of DAz-2-labeled E. coli ThrRS and western blot analysis DAz-2-labeled ThrRS was conjugated to biotin via Staudinger ligation with 0.25 mM phosphine-PEG3-biotin (Thermo Scientific) at 37°C for 2 h. Biotinylation reactions were terminated by the addition of 1 ml of cold acetone and were kept in −80°C for 20 min. .. Protein pellet was washed once by 200 μl acetone then resuspended in 2x sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) loading dye.

    Protease Inhibitor:

    Article Title: Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis
    Article Snippet: For western blot analysis, the PC-3 tumor cells were washed twice with DPBS and were lysed using RIPA buffer with 1% protease inhibitor. .. The total protein of each lysate was quantified by BCA assay and the lysates were incubated with 500 nM of phosphine–PEG3 -biotin (Pierce, Rockford, IL, USA) for 6 h at 37 °C.

    Article Title: Cell Labeling and Tracking Method without Distorted Signals by Phagocytosis of Macrophages
    Article Snippet: They were pelleted by centrifugation at 3,000×g for 5 min, and the cell pellets were resuspended in 500 µL of lysis buffer (1% SDS, 100 mM Tris-HCl, pH 7.4) containing protease inhibitor cocktail (Complete, EDTA-free, Roche, NSW, Australia) and lysed with a probe-type sonifier at 4 °C. .. Then, 20 µL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3 -biotin (2 µL, 5 mM in DPBS) (Pierce) for 6 h at 37 o C. Loading buffer was added to each sample, and samples were loaded onto 10% SPS-PAGE gel after heating at 95 o C. Proteins were transferred to Hybond P membrane (Amercham, St. Albans, UK), and the membrane was blocked with 5% bovine serum albumin in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH 7.4) for 2 h. The membrane was then incubated with streptavidin-horseradish peroxidase (diluted 1:2,000 in TBST) (Pierce) overnight at 4 o C. The membrane was rinsed three times with TBST and developed by using ECL Western Blotting Substrate (Pierce).

    Article Title: Physiological Effects of Ac4ManNAz and Optimization of Metabolic Labeling for Cell Tracking
    Article Snippet: They were pelleted by centrifugation at 3,000×g for 5 min, and the cell pellets were resuspended in 500 μL of lysis buffer (1% SDS, 100 mM Tris-HCl, pH 7.4) containing protease inhibitor cocktail (Complete, EDTA-free, Roche, NSW, Australia) and lysed with a probe-type sonifier at 4 °C. .. Then, 20 μL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3-biotin (2 μL, 5 mM in DPBS) (Pierce) for 6 h at 37 °C.

    Article Title: Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis
    Article Snippet: Streptavidin-horseradish peroxidase (streptavidin-HRP) and Phosphine-PEG3 -biotin were purchased from Thermo Fisher Scientific Inc. (Rockford, USA). .. Dimethylformaide (DMF), trifluoroacetic acid (TFA), dimethyl sulfoxide (DMSO), 4-nitrophenyl chloroformate (4-NPC), N,N-dimethylpyridin-4-amine (DMAP), doxorubicin hydrochloride (DOX), Annexin-V assay kit and protease inhibitor cocktail were purchased from Sigma-Aldrich (St. Louis, USA).

    Bicinchoninic Acid Protein Assay:

    Article Title: Cell Labeling and Tracking Method without Distorted Signals by Phagocytosis of Macrophages
    Article Snippet: Final soluble protein concentrations were determined by bicinchoninic acid protein assay (Pierce, Rockford, IL, USA) to be 5 mg/mL. .. Then, 20 µL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3 -biotin (2 µL, 5 mM in DPBS) (Pierce) for 6 h at 37 o C. Loading buffer was added to each sample, and samples were loaded onto 10% SPS-PAGE gel after heating at 95 o C. Proteins were transferred to Hybond P membrane (Amercham, St. Albans, UK), and the membrane was blocked with 5% bovine serum albumin in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH 7.4) for 2 h. The membrane was then incubated with streptavidin-horseradish peroxidase (diluted 1:2,000 in TBST) (Pierce) overnight at 4 o C. The membrane was rinsed three times with TBST and developed by using ECL Western Blotting Substrate (Pierce).

    Article Title: Physiological Effects of Ac4ManNAz and Optimization of Metabolic Labeling for Cell Tracking
    Article Snippet: Final soluble protein concentrations were determined by bicinchoninic acid protein assay (Pierce, Rockford, IL, USA) to be 5 mg/mL. .. Then, 20 μL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3-biotin (2 μL, 5 mM in DPBS) (Pierce) for 6 h at 37 °C.

    Labeling:

    Article Title: Bioorthogonal labeling cell-surface proteins expressed in pancreatic cancer cells to identify potential diagnostic/therapeutic biomarkers
    Article Snippet: Paragraph title: Metabolic labeling cells ... After 72 h, the cells were washed twice with phosphate-buffered saline (PBS) then treated with culture medium containing 200 µM of either EZ-Link Phosphine-PEG3 -Biotin (Thermo Fisher Scientific, 88901) or DBCO-S-S-PEG3-Biotin (Click Chemistry Tools, A112) conjugate suspended in DMSO to selectively tag glycoproteins on the surface of live cells.

    Article Title: A molecular brake controls the magnitude of long-term potentiation
    Article Snippet: Paragraph title: Metabolic labeling of synaptoneurosomes and immunoprecipitation ... Cortical synaptoneurosomes were pre-incubated with 500 µM AHA in the absence and presence of 1 µM rapamycin for 20 min, and then treated with 100 ng/ml of BDNF for 60 min. After washing out the treatments, samples were incubated with phosphine-PEG3-Biotin (Thermo) for 4 h at 37 °C, to conjugate biotin to AHA containing proteins.

    Article Title: The non-coding RNA BC1 regulates experience-dependent structural plasticity and learning
    Article Snippet: Paragraph title: Metabolic labeling of synaptoneurosomes ... Cortical synaptoneurosomes were pre-incubated with 500 µM AHA in Hepes-Krebs buffer for 50 min at 37 °C, and then treated with 100 µM DHPG for 10 min. After washing out the treatments, samples were solubilized in lysis buffer containing PIC and incubated with 500 µM phosphine-PEG3-Biotin (Thermo) for 4 h at 37 °C to conjugate biotin to AHA-containing proteins.

    Concentration Assay:

    Article Title: Bioorthogonal labeling cell-surface proteins expressed in pancreatic cancer cells to identify potential diagnostic/therapeutic biomarkers
    Article Snippet: The following day, 10 mM N -azidoacetylmannosamine-tetraacylated prepared in DMSO (Ac4 ManNAz, Thermo Fisher Scientific, 88904) was added to 2 dishes of cells to yield a final concentration of 50 µM azido sugar. .. After 72 h, the cells were washed twice with phosphate-buffered saline (PBS) then treated with culture medium containing 200 µM of either EZ-Link Phosphine-PEG3 -Biotin (Thermo Fisher Scientific, 88901) or DBCO-S-S-PEG3-Biotin (Click Chemistry Tools, A112) conjugate suspended in DMSO to selectively tag glycoproteins on the surface of live cells.

    Immunoprecipitation:

    Article Title: A molecular brake controls the magnitude of long-term potentiation
    Article Snippet: Paragraph title: Metabolic labeling of synaptoneurosomes and immunoprecipitation ... Cortical synaptoneurosomes were pre-incubated with 500 µM AHA in the absence and presence of 1 µM rapamycin for 20 min, and then treated with 100 ng/ml of BDNF for 60 min. After washing out the treatments, samples were incubated with phosphine-PEG3-Biotin (Thermo) for 4 h at 37 °C, to conjugate biotin to AHA containing proteins.

    Incubation:

    Article Title: A molecular brake controls the magnitude of long-term potentiation
    Article Snippet: .. Cortical synaptoneurosomes were pre-incubated with 500 µM AHA in the absence and presence of 1 µM rapamycin for 20 min, and then treated with 100 ng/ml of BDNF for 60 min. After washing out the treatments, samples were incubated with phosphine-PEG3-Biotin (Thermo) for 4 h at 37 °C, to conjugate biotin to AHA containing proteins. .. Samples were then eluted on Zeba Spin desalting columns (7K MWCO, Thermo) to remove the excess of phosphine-PEG3-Biotin.

    Article Title: Mechanism of oxidant-induced mistranslation by threonyl-tRNA synthetase
    Article Snippet: Biotinylation of DAz-2-labeled E. coli ThrRS and western blot analysis DAz-2-labeled ThrRS was conjugated to biotin via Staudinger ligation with 0.25 mM phosphine-PEG3-biotin (Thermo Scientific) at 37°C for 2 h. Biotinylation reactions were terminated by the addition of 1 ml of cold acetone and were kept in −80°C for 20 min. .. The membrane was blocked in 3% bovine serum albumin (Fisher) in Tris-buffered saline Tween-20 (TBST; 25 mM Tris/Tris-HCl, 137 mM NaCl, 2.7 mM KCl and 0.05% Tween-20) at room temperature for 1 h. Blocked membrane was then incubated with 1:50 000 dilution of streptavidin-horseradish peroxidase (HRP) (GE Healthcare) at room temperature for 1 h, washed three times with 50 ml of TBST and developed with Clarity Western enhanced chemiluminescence (ECL) Substrate (Bio-Rad).

    Article Title: Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis
    Article Snippet: .. The total protein of each lysate was quantified by BCA assay and the lysates were incubated with 500 nM of phosphine–PEG3 -biotin (Pierce, Rockford, IL, USA) for 6 h at 37 °C. .. The proteins from each sample were mixed with 1 x sodium dodecyl sulfate (SDS) gel-loading buffer and boiled for 5 min. 20 μg of proteins were separated by 10% SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membranes.

    Article Title: Cell Labeling and Tracking Method without Distorted Signals by Phagocytosis of Macrophages
    Article Snippet: .. Then, 20 µL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3 -biotin (2 µL, 5 mM in DPBS) (Pierce) for 6 h at 37 o C. Loading buffer was added to each sample, and samples were loaded onto 10% SPS-PAGE gel after heating at 95 o C. Proteins were transferred to Hybond P membrane (Amercham, St. Albans, UK), and the membrane was blocked with 5% bovine serum albumin in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH 7.4) for 2 h. The membrane was then incubated with streptavidin-horseradish peroxidase (diluted 1:2,000 in TBST) (Pierce) overnight at 4 o C. The membrane was rinsed three times with TBST and developed by using ECL Western Blotting Substrate (Pierce). .. Cellular imaging to determine the generated azide groups A549 cells were seeded onto 35-mm glass-bottom dishes at a density of 3×104 cells in 2 mL of growth media with Ac4 ManNAz (50 µM, final concentration).

    Article Title: Physiological Effects of Ac4ManNAz and Optimization of Metabolic Labeling for Cell Tracking
    Article Snippet: .. Then, 20 μL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3-biotin (2 μL, 5 mM in DPBS) (Pierce) for 6 h at 37 °C. ..

    Article Title: The non-coding RNA BC1 regulates experience-dependent structural plasticity and learning
    Article Snippet: .. Cortical synaptoneurosomes were pre-incubated with 500 µM AHA in Hepes-Krebs buffer for 50 min at 37 °C, and then treated with 100 µM DHPG for 10 min. After washing out the treatments, samples were solubilized in lysis buffer containing PIC and incubated with 500 µM phosphine-PEG3-Biotin (Thermo) for 4 h at 37 °C to conjugate biotin to AHA-containing proteins. .. Samples were then eluted on Zeba Spin desalting columns (7K MWCO, Thermo) to remove the excess of phosphine-PEG3-Biotin.

    Article Title: The Metabolic Chemical Reporter 6‑Azido-6-deoxy-glucose Further Reveals the Substrate Promiscuity of O‑GlcNAc Transferase and Catalyzes the Discovery of Intracellular Protein Modification by O‑Glucose
    Article Snippet: .. The mixture was sonicated in a bath sonicator to ensure complete dissolution, and then 73 μ L of H2 O was added, and 2 μ L of 10 mM EZ-Link Phosphine-PEG3 -Biotin (10 mM in DMSO, Thermo Scientific) was added and then incubated for 3 h at 37 °C and quenched with chloroform/ methanol precipitation. ..

    Cell Culture:

    Article Title: Physiological Effects of Ac4ManNAz and Optimization of Metabolic Labeling for Cell Tracking
    Article Snippet: Western blot analysis A549 cells were seeded onto 100×20 mm cell culture plates at a density of 2×106 cells per plate in 10 mL of media with no sugar or Ac4ManNAz (10, 20 and 50 μM). .. Then, 20 μL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3-biotin (2 μL, 5 mM in DPBS) (Pierce) for 6 h at 37 °C.

    Protein Concentration:

    Article Title: The Metabolic Chemical Reporter 6‑Azido-6-deoxy-glucose Further Reveals the Substrate Promiscuity of O‑GlcNAc Transferase and Catalyzes the Discovery of Intracellular Protein Modification by O‑Glucose
    Article Snippet: The mixture was sonicated in a bath sonicator to ensure complete dissolution, and then 73 μ L of H2 O was added, and 2 μ L of 10 mM EZ-Link Phosphine-PEG3 -Biotin (10 mM in DMSO, Thermo Scientific) was added and then incubated for 3 h at 37 °C and quenched with chloroform/ methanol precipitation. .. The mixture was sonicated in a bath sonicator to ensure complete dissolution, and then 73 μ L of H2 O was added, and 2 μ L of 10 mM EZ-Link Phosphine-PEG3 -Biotin (10 mM in DMSO, Thermo Scientific) was added and then incubated for 3 h at 37 °C and quenched with chloroform/ methanol precipitation.

    Annexin V Assay:

    Article Title: Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis
    Article Snippet: Streptavidin-horseradish peroxidase (streptavidin-HRP) and Phosphine-PEG3 -biotin were purchased from Thermo Fisher Scientific Inc. (Rockford, USA). .. Dimethylformaide (DMF), trifluoroacetic acid (TFA), dimethyl sulfoxide (DMSO), 4-nitrophenyl chloroformate (4-NPC), N,N-dimethylpyridin-4-amine (DMAP), doxorubicin hydrochloride (DOX), Annexin-V assay kit and protease inhibitor cocktail were purchased from Sigma-Aldrich (St. Louis, USA).

    Western Blot:

    Article Title: Mechanism of oxidant-induced mistranslation by threonyl-tRNA synthetase
    Article Snippet: .. Biotinylation of DAz-2-labeled E. coli ThrRS and western blot analysis DAz-2-labeled ThrRS was conjugated to biotin via Staudinger ligation with 0.25 mM phosphine-PEG3-biotin (Thermo Scientific) at 37°C for 2 h. Biotinylation reactions were terminated by the addition of 1 ml of cold acetone and were kept in −80°C for 20 min. .. Protein pellet was washed once by 200 μl acetone then resuspended in 2x sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) loading dye.

    Article Title: Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis
    Article Snippet: Paragraph title: Western blot analysis for azido (N3 ) groups-containing sialic acid in PC-3 tumor cells ... The total protein of each lysate was quantified by BCA assay and the lysates were incubated with 500 nM of phosphine–PEG3 -biotin (Pierce, Rockford, IL, USA) for 6 h at 37 °C.

    Article Title: Cell Labeling and Tracking Method without Distorted Signals by Phagocytosis of Macrophages
    Article Snippet: .. Then, 20 µL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3 -biotin (2 µL, 5 mM in DPBS) (Pierce) for 6 h at 37 o C. Loading buffer was added to each sample, and samples were loaded onto 10% SPS-PAGE gel after heating at 95 o C. Proteins were transferred to Hybond P membrane (Amercham, St. Albans, UK), and the membrane was blocked with 5% bovine serum albumin in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH 7.4) for 2 h. The membrane was then incubated with streptavidin-horseradish peroxidase (diluted 1:2,000 in TBST) (Pierce) overnight at 4 o C. The membrane was rinsed three times with TBST and developed by using ECL Western Blotting Substrate (Pierce). .. Cellular imaging to determine the generated azide groups A549 cells were seeded onto 35-mm glass-bottom dishes at a density of 3×104 cells in 2 mL of growth media with Ac4 ManNAz (50 µM, final concentration).

    Article Title: Physiological Effects of Ac4ManNAz and Optimization of Metabolic Labeling for Cell Tracking
    Article Snippet: Paragraph title: Western blot analysis ... Then, 20 μL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3-biotin (2 μL, 5 mM in DPBS) (Pierce) for 6 h at 37 °C.

    Article Title: Selective in vivo metabolic cell-labeling-mediated cancer targeting
    Article Snippet: .. EZ-Link phosphine–PEG3 –biotin, streptavidin–horseradish peroxidase (HRP), and Pierce ECL western blotting substrate were purchased from Thermo Fisher Scientific (Waltham, MA, USA). ..

    Lysis:

    Article Title: Cell Labeling and Tracking Method without Distorted Signals by Phagocytosis of Macrophages
    Article Snippet: They were pelleted by centrifugation at 3,000×g for 5 min, and the cell pellets were resuspended in 500 µL of lysis buffer (1% SDS, 100 mM Tris-HCl, pH 7.4) containing protease inhibitor cocktail (Complete, EDTA-free, Roche, NSW, Australia) and lysed with a probe-type sonifier at 4 °C. .. Then, 20 µL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3 -biotin (2 µL, 5 mM in DPBS) (Pierce) for 6 h at 37 o C. Loading buffer was added to each sample, and samples were loaded onto 10% SPS-PAGE gel after heating at 95 o C. Proteins were transferred to Hybond P membrane (Amercham, St. Albans, UK), and the membrane was blocked with 5% bovine serum albumin in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH 7.4) for 2 h. The membrane was then incubated with streptavidin-horseradish peroxidase (diluted 1:2,000 in TBST) (Pierce) overnight at 4 o C. The membrane was rinsed three times with TBST and developed by using ECL Western Blotting Substrate (Pierce).

    Article Title: Physiological Effects of Ac4ManNAz and Optimization of Metabolic Labeling for Cell Tracking
    Article Snippet: They were pelleted by centrifugation at 3,000×g for 5 min, and the cell pellets were resuspended in 500 μL of lysis buffer (1% SDS, 100 mM Tris-HCl, pH 7.4) containing protease inhibitor cocktail (Complete, EDTA-free, Roche, NSW, Australia) and lysed with a probe-type sonifier at 4 °C. .. Then, 20 μL of the lysate (5 mg/mL protein) was incubated with phosphine-PEG3-biotin (2 μL, 5 mM in DPBS) (Pierce) for 6 h at 37 °C.

    Article Title: The non-coding RNA BC1 regulates experience-dependent structural plasticity and learning
    Article Snippet: .. Cortical synaptoneurosomes were pre-incubated with 500 µM AHA in Hepes-Krebs buffer for 50 min at 37 °C, and then treated with 100 µM DHPG for 10 min. After washing out the treatments, samples were solubilized in lysis buffer containing PIC and incubated with 500 µM phosphine-PEG3-Biotin (Thermo) for 4 h at 37 °C to conjugate biotin to AHA-containing proteins. .. Samples were then eluted on Zeba Spin desalting columns (7K MWCO, Thermo) to remove the excess of phosphine-PEG3-Biotin.

    Recombinant:

    Article Title: Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis
    Article Snippet: Streptavidin-horseradish peroxidase (streptavidin-HRP) and Phosphine-PEG3 -biotin were purchased from Thermo Fisher Scientific Inc. (Rockford, USA). .. Recombinant Human TRAIL/TNFSF10 protein, recombinant Human Cas-3 protein, Human/Mouse active Cas-3 antibody, Human/Mouse Cas-8 antibody, goat polyclonal GAPDH antibody, recombinant Human Cas-1, recombinant Human Cas-8, recombinant Human Cas-7, recombinant Human Cathepsin B and recombinant Human MMP-9 and N-benzyloxycarbonyl-Asp(OMe)-Glu(OMe)-Val-Asp(OMe)-fluoromethylketone (z-DEVD-FMK, Cas-3/-7 inhibitor) were purchased from R & D System (Minneapolis, USA).

    BIA-KA:

    Article Title: Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis
    Article Snippet: .. The total protein of each lysate was quantified by BCA assay and the lysates were incubated with 500 nM of phosphine–PEG3 -biotin (Pierce, Rockford, IL, USA) for 6 h at 37 °C. .. The proteins from each sample were mixed with 1 x sodium dodecyl sulfate (SDS) gel-loading buffer and boiled for 5 min. 20 μg of proteins were separated by 10% SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membranes.

    Article Title: The Metabolic Chemical Reporter 6‑Azido-6-deoxy-glucose Further Reveals the Substrate Promiscuity of O‑GlcNAc Transferase and Catalyzes the Discovery of Intracellular Protein Modification by O‑Glucose
    Article Snippet: The mixture was sonicated in a bath sonicator to ensure complete dissolution, and then 73 μ L of H2 O was added, and 2 μ L of 10 mM EZ-Link Phosphine-PEG3 -Biotin (10 mM in DMSO, Thermo Scientific) was added and then incubated for 3 h at 37 °C and quenched with chloroform/ methanol precipitation. .. The mixture was sonicated in a bath sonicator to ensure complete dissolution, and then 73 μ L of H2 O was added, and 2 μ L of 10 mM EZ-Link Phosphine-PEG3 -Biotin (10 mM in DMSO, Thermo Scientific) was added and then incubated for 3 h at 37 °C and quenched with chloroform/ methanol precipitation.

    SDS Page:

    Article Title: Mechanism of oxidant-induced mistranslation by threonyl-tRNA synthetase
    Article Snippet: Biotinylation of DAz-2-labeled E. coli ThrRS and western blot analysis DAz-2-labeled ThrRS was conjugated to biotin via Staudinger ligation with 0.25 mM phosphine-PEG3-biotin (Thermo Scientific) at 37°C for 2 h. Biotinylation reactions were terminated by the addition of 1 ml of cold acetone and were kept in −80°C for 20 min. .. Protein pellet was washed once by 200 μl acetone then resuspended in 2x sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) loading dye.

    Article Title: The Metabolic Chemical Reporter 6‑Azido-6-deoxy-glucose Further Reveals the Substrate Promiscuity of O‑GlcNAc Transferase and Catalyzes the Discovery of Intracellular Protein Modification by O‑Glucose
    Article Snippet: The mixture was sonicated in a bath sonicator to ensure complete dissolution, and then 73 μ L of H2 O was added, and 2 μ L of 10 mM EZ-Link Phosphine-PEG3 -Biotin (10 mM in DMSO, Thermo Scientific) was added and then incubated for 3 h at 37 °C and quenched with chloroform/ methanol precipitation. .. The samples were boiled for 5 min at 97 °C, and 10 or 5 μ g of protein was then loaded per lane for SDS-PAGE separation.

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  • 88
    Thermo Fisher ez link phosphine peg3 biotin
    Ez Link Phosphine Peg3 Biotin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ez link phosphine peg3 biotin/product/Thermo Fisher
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ez link phosphine peg3 biotin - by Bioz Stars, 2020-01
    88/100 stars
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