streptavidin agarose resin  (Millipore)


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    Structured Review

    Millipore streptavidin agarose resin
    N63Y mutant CVB3 has a growth defect in cell culture and reduced glycan-mediated cell attachment. Single-cycle assays of viral replication in HeLa (A) and Huh7 (B) cells were performed. Infections with WT or N63Y mutant CVB3 were performed at an MOI of 0.1. Viral titers were determined by plaque assay with HeLa cells. n = 3. (C) Cell attachment of 35 S-labeled WT or N63Y mutant CVB3. Virus was incubated with cells at 4°C for 40 min. Cells were washed and trypsinized, and cell-associated 35 S was quantified. (D) 35 S-labeled WT or N63Y mutant CVB3 was incubated with CHO cells (CHO-K1, pgsA745, pgsD677, and pgsB761) that vary in GAG expression. Plus and minus signs indicate the relative levels of GAGs on the cell surface. (E) Effect of heparinase treatment on CVB3 cell attachment. Huh7 cells were treated with or without heparinase I for 90 min prior to quantification of 35 S CVB3 attachment. n = 7. (F) Heparin-agarose pulldown assay. 35 S-labeled WT or N63Y mutant CVB3 was incubated with heparin-agarose resin or <t>streptavidin-agarose</t> resin (control). Resin was washed, and bound 35 S-labeled CVB3 was quantified. n = 3. *, P
    Streptavidin Agarose Resin, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Emergence of a Large-Plaque Variant in Mice Infected with Coxsackievirus B3"

    Article Title: Emergence of a Large-Plaque Variant in Mice Infected with Coxsackievirus B3

    Journal: mBio

    doi: 10.1128/mBio.00119-16

    N63Y mutant CVB3 has a growth defect in cell culture and reduced glycan-mediated cell attachment. Single-cycle assays of viral replication in HeLa (A) and Huh7 (B) cells were performed. Infections with WT or N63Y mutant CVB3 were performed at an MOI of 0.1. Viral titers were determined by plaque assay with HeLa cells. n = 3. (C) Cell attachment of 35 S-labeled WT or N63Y mutant CVB3. Virus was incubated with cells at 4°C for 40 min. Cells were washed and trypsinized, and cell-associated 35 S was quantified. (D) 35 S-labeled WT or N63Y mutant CVB3 was incubated with CHO cells (CHO-K1, pgsA745, pgsD677, and pgsB761) that vary in GAG expression. Plus and minus signs indicate the relative levels of GAGs on the cell surface. (E) Effect of heparinase treatment on CVB3 cell attachment. Huh7 cells were treated with or without heparinase I for 90 min prior to quantification of 35 S CVB3 attachment. n = 7. (F) Heparin-agarose pulldown assay. 35 S-labeled WT or N63Y mutant CVB3 was incubated with heparin-agarose resin or streptavidin-agarose resin (control). Resin was washed, and bound 35 S-labeled CVB3 was quantified. n = 3. *, P
    Figure Legend Snippet: N63Y mutant CVB3 has a growth defect in cell culture and reduced glycan-mediated cell attachment. Single-cycle assays of viral replication in HeLa (A) and Huh7 (B) cells were performed. Infections with WT or N63Y mutant CVB3 were performed at an MOI of 0.1. Viral titers were determined by plaque assay with HeLa cells. n = 3. (C) Cell attachment of 35 S-labeled WT or N63Y mutant CVB3. Virus was incubated with cells at 4°C for 40 min. Cells were washed and trypsinized, and cell-associated 35 S was quantified. (D) 35 S-labeled WT or N63Y mutant CVB3 was incubated with CHO cells (CHO-K1, pgsA745, pgsD677, and pgsB761) that vary in GAG expression. Plus and minus signs indicate the relative levels of GAGs on the cell surface. (E) Effect of heparinase treatment on CVB3 cell attachment. Huh7 cells were treated with or without heparinase I for 90 min prior to quantification of 35 S CVB3 attachment. n = 7. (F) Heparin-agarose pulldown assay. 35 S-labeled WT or N63Y mutant CVB3 was incubated with heparin-agarose resin or streptavidin-agarose resin (control). Resin was washed, and bound 35 S-labeled CVB3 was quantified. n = 3. *, P

    Techniques Used: Mutagenesis, Cell Culture, Cell Attachment Assay, Plaque Assay, Labeling, Incubation, Expressing

    2) Product Images from "Enhanced Functional Potential of Nucleic Acid Aptamer Libraries Patterned to Increase Secondary Structure"

    Article Title: Enhanced Functional Potential of Nucleic Acid Aptamer Libraries Patterned to Increase Secondary Structure

    Journal: Journal of the American Chemical Society

    doi: 10.1021/ja103023m

    Streptavidin binding motif analysis. (a) The binding activities of point mutants (listed in parentheses) reveal a consensus set of binding motifs. (b) The most likely frame for this consensus sequence in the R*Y* library is shown.
    Figure Legend Snippet: Streptavidin binding motif analysis. (a) The binding activities of point mutants (listed in parentheses) reveal a consensus set of binding motifs. (b) The most likely frame for this consensus sequence in the R*Y* library is shown.

    Techniques Used: Binding Assay, Sequencing

    Binding activity of pools over 9 to 10 rounds of selections for binding (a) streptavidin, (b) IgE, and (c) VEGF.
    Figure Legend Snippet: Binding activity of pools over 9 to 10 rounds of selections for binding (a) streptavidin, (b) IgE, and (c) VEGF.

    Techniques Used: Binding Assay, Activity Assay

    3) Product Images from "Function of glutathione peroxidases in legume root nodules"

    Article Title: Function of glutathione peroxidases in legume root nodules

    Journal: Journal of Experimental Botany

    doi: 10.1093/jxb/erv066

    Nitrosylation of LjGpx1 and LjGpx3. (A) Immunoblot showing nitrosylation of purified LjGpx1 and LjGpx3. Recombinant proteins were treated with (-) 1mM GSSG (control) or with (+) 1mM GSNO, subjected to the biotin-switch, and immunoblotted with an anti-biotin antibody. (B) Immunoblot showing nitrosylation of LjGpx1 and LjGpx3 in nodule extracts from plants treated with 5mM GSSG (-) or with 5mM GSNO (+). Biotinylated proteins were affinity purified using streptavidin-agarose and immunoblotted with the LjGpx1 and LjGpx3 antibodies. (C) Effect of GSNO-mediated nitrosylation on LjGpx activities measured using the NADPH-coupled assay with poplar Trx h1 and H 2 O 2 as substrates. Recombinant LjGpx1 and LjGpx3 were treated with 1mM GSSG (control), 1mM GSNO, or 5mM GSNO for 1h at 37ºC. Values are means ± SE of 6–8 replicates. Means marked with an asterisk differ significantly from control at P
    Figure Legend Snippet: Nitrosylation of LjGpx1 and LjGpx3. (A) Immunoblot showing nitrosylation of purified LjGpx1 and LjGpx3. Recombinant proteins were treated with (-) 1mM GSSG (control) or with (+) 1mM GSNO, subjected to the biotin-switch, and immunoblotted with an anti-biotin antibody. (B) Immunoblot showing nitrosylation of LjGpx1 and LjGpx3 in nodule extracts from plants treated with 5mM GSSG (-) or with 5mM GSNO (+). Biotinylated proteins were affinity purified using streptavidin-agarose and immunoblotted with the LjGpx1 and LjGpx3 antibodies. (C) Effect of GSNO-mediated nitrosylation on LjGpx activities measured using the NADPH-coupled assay with poplar Trx h1 and H 2 O 2 as substrates. Recombinant LjGpx1 and LjGpx3 were treated with 1mM GSSG (control), 1mM GSNO, or 5mM GSNO for 1h at 37ºC. Values are means ± SE of 6–8 replicates. Means marked with an asterisk differ significantly from control at P

    Techniques Used: Purification, Recombinant, Affinity Purification

    Related Articles

    Centrifugation:

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: .. After centrifugation, supernatants were incubated with streptavidin agarose resin (Millipore) for 5 h at 4 °C. .. Beads were washed with lysis buffer and TEV buffer (50 mM Tris-HCl pH 7.5, 1 mM ATP, and 10% glycerol).

    Article Title: Distinct tRNA recognition strategies used by a homologous family of editing domains prevent mistranslation
    Article Snippet: .. Biotinylated dFx was removed by incubating the reaction mixture with 150 μl of streptavidin agarose resin (Novagen) for ∼15 min at room temperature followed by centrifugation in a table top centrifuge at 4°C for 2 min at 2000×g . ..

    Article Title: Insights into Brain Glycogen Metabolism
    Article Snippet: Biotinylated thrombin was removed from the mixture by the addition of streptavidin-agarose resin (16 μl/unit thrombin, Sigma-Aldrich) for 30 min at 4 °C with agitation. .. Biotinylated thrombin was removed from the mixture by the addition of streptavidin-agarose resin (16 μl/unit thrombin, Sigma-Aldrich) for 30 min at 4 °C with agitation.

    Article Title: Function of glutathione peroxidases in legume root nodules
    Article Snippet: Dry pellets were resuspended in binding buffer consisting of 25mM HEPES (pH 7.7), 1mM EDTA, 100mM NaCl, 0.8% Triton X-100, and 50 μl of streptavidin-agarose resin (Sigma). .. After centrifugation, proteins were separated on 15% SDS gels and transferred to membranes for immunoblot analysis with LjGpx antibodies.

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: .. After centrifugation, supernatants were incubated with RNA aptamers for 10 min, and then with streptavidin agarose resin (Millipore) for 5 h at 4 °C. .. Beads were washed with lysis buffer and boiled in SDS sample buffer.

    Article Title: Emergence of a Large-Plaque Variant in Mice Infected with Coxsackievirus B3
    Article Snippet: 35 S-labeled CVB3 (5 × 106 PFU/5,000 cpm) was mixed with heparin agarose resin (Sigma Aldrich) or streptavidin agarose resin (Sigma Aldrich). .. Following incubation, resin was pelleted by centrifugation at 2,600 × g for 2 min. Resin was washed three times with PBS+ (PBS supplemented with 100 μg/ml CaCl2 and 100 μg/ml MgCl2 ), and resin-associated radioactivity was quantified by scintillation counting.

    Article Title: Ancestral AlaX Editing Enzymes for Control of Genetic Code Fidelity Are Not tRNA-specific *
    Article Snippet: .. Biotinylated dFx was removed by incubating the reaction mixture with 150 μl of streptavidin-agarose resin (Novagen) for ∼15 min at room temperature, followed by centrifugation in a tabletop centrifuge at 4 °C for 2 min at 2000 × g . ..

    Synthesized:

    Article Title: The RNA Binding Protein hnRNP Q Modulates the Utilization of Exon 7 in the Survival Motor Neuron 2 (SMN2) Gene ▿) Gene ▿ †
    Article Snippet: Biotinylated RNA oligonucleotides corresponding to SMN1 and SMN2 and containing the 3′ 13 residues of intron 6 and the 5′ 22 residues of exon 7 were synthesized by Dharmacon. .. To identify HeLa cell proteins that associate with SMN exon 7, 1.2 μg of biotinylated RNA oligonucleotide was conjugated with 20 μl of streptavidin-agarose resin (Sigma).

    Incubation:

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: .. After centrifugation, supernatants were incubated with streptavidin agarose resin (Millipore) for 5 h at 4 °C. .. Beads were washed with lysis buffer and TEV buffer (50 mM Tris-HCl pH 7.5, 1 mM ATP, and 10% glycerol).

    Article Title: Nucleotide Excision Repair Is Associated with the Replisome and Its Efficiency Depends on a Direct Interaction between XPA and PCNA
    Article Snippet: Cells were incubated in click reaction buffer for 1–2 h at a concentration of 1×108 cells per 5 ml of click reaction buffer (2 mM CuSO4 , 10 µM biotin-azide, 10 mM Sodium ascorbate). .. Streptavidin–agarose resin (100 µl resin per 2×108 cells, Novagen) was washed twice in lysis buffer and once in PBS.

    Article Title: Insights into Brain Glycogen Metabolism
    Article Snippet: Briefly, 20 mg of 6xHis-bGP was incubated with biotinylated-thrombin (0.5 units thrombin/mg protein, purchased from Merck-Millipore) for 3 h on ice. .. Biotinylated thrombin was removed from the mixture by the addition of streptavidin-agarose resin (16 μl/unit thrombin, Sigma-Aldrich) for 30 min at 4 °C with agitation.

    Article Title: Function of glutathione peroxidases in legume root nodules
    Article Snippet: Dry pellets were resuspended in binding buffer consisting of 25mM HEPES (pH 7.7), 1mM EDTA, 100mM NaCl, 0.8% Triton X-100, and 50 μl of streptavidin-agarose resin (Sigma). .. Samples were incubated overnight at 4ºC and then the agarose beads were washed ten times with a buffer comprising 25mM HEPES (pH 7.7), 1mM EDTA, 600mM NaCl, and 0.8% Triton X-100.

    Article Title: Latent Membrane Protein 2A of Epstein-Barr Virus Binds WW Domain E3 Protein-Ubiquitin Ligases That Ubiquitinate B-Cell Tyrosine Kinases
    Article Snippet: For each affinity reagent, a 100-μl-bed volume of streptavidin-agarose resin (Sigma) capable of binding 10 nmol of biotin was washed with 20 mM sodium phosphate buffer (pH 7.0) containing 150 mM NaCl (PBS) and then resuspended in a twofold-molar excess of biotinylated peptide. .. Following a 15-min incubation and mixing at room temperature, the resin was washed three times with excess PBS.

    Article Title: Cryo-EM structure of substrate-bound human telomerase holoenzyme
    Article Snippet: .. The prepared whole-cell extract (from 120 plates of adherent cells or 6L of suspension cells) was incubated with streptavidin agarose resin (Sigma-Aldrich) pre-bound to a 5’-biotinylated oligonucleotide at room temperature for 3 hours. ..

    Article Title: The RNA Binding Protein hnRNP Q Modulates the Utilization of Exon 7 in the Survival Motor Neuron 2 (SMN2) Gene ▿) Gene ▿ †
    Article Snippet: Subsequently, a 70-μl reaction mixture containing 50 μl of precleaned nuclear extract and 5 μg of biotinylated RNA was incubated under splicing conditions (0.5 mM ATP, 20 mM creatine phosphate, 2.4 mM MgCl, and 20 units of RNasin; Promega) at 30°C for 30 min, followed by a further 3-h incubation with 50 μl of streptavidin beads at 4°C. .. To identify HeLa cell proteins that associate with SMN exon 7, 1.2 μg of biotinylated RNA oligonucleotide was conjugated with 20 μl of streptavidin-agarose resin (Sigma).

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: .. After centrifugation, supernatants were incubated with RNA aptamers for 10 min, and then with streptavidin agarose resin (Millipore) for 5 h at 4 °C. .. Beads were washed with lysis buffer and boiled in SDS sample buffer.

    Article Title: Emergence of a Large-Plaque Variant in Mice Infected with Coxsackievirus B3
    Article Snippet: 35 S-labeled CVB3 (5 × 106 PFU/5,000 cpm) was mixed with heparin agarose resin (Sigma Aldrich) or streptavidin agarose resin (Sigma Aldrich). .. Following incubation, resin was pelleted by centrifugation at 2,600 × g for 2 min. Resin was washed three times with PBS+ (PBS supplemented with 100 μg/ml CaCl2 and 100 μg/ml MgCl2 ), and resin-associated radioactivity was quantified by scintillation counting.

    Article Title: Enhanced Functional Potential of Nucleic Acid Aptamer Libraries Patterned to Increase Secondary Structure
    Article Snippet: .. The DNA was added to 40 μL of streptavidin-agarose resin (Novagen) and incubated with end-over-end rotation for 2 h at 25 °C. .. The DNA and resin were transferred to a 5.0 μm Amicon Ultrafree-MC spin filter (Millipore) and centrifuged to separate the supernatant from the resin.

    Article Title: Correlated S-palmitoylation profiling of Snail-induced epithelial to mesenchymal transition
    Article Snippet: Both +NH2 OH and −NH2 OH samples were chloroform-methanol extracted, washed twice with methanol, resuspended in 4% SDS/PBS, and incubated with 200 μM biotin-PEG-maleimide (Click Chemistry Tools) for 90 minutes. .. Samples were then diluted to 0.5 mg/mL protein and 0.25% SDS in DPBS, and combined with 50 μL of streptavidin-agarose resin (Millipore) rotating for 90 minutes at room temperature.

    Activity Assay:

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: After centrifugation, supernatants were incubated with streptavidin agarose resin (Millipore) for 5 h at 4 °C. .. To inhibit the deubiquitinating activity of proteasomes, ubiquitin-vinylmethylester (Ub-vme; LifeSensors) was added as previously described .

    Article Title: Single-molecule imaging of telomerase reverse transcriptase in human telomerase holoenzyme and minimal RNP complexes
    Article Snippet: .. Enrichment of complexes by tagged TERT, tagged TPP1, or hTR template for activity assays HEK 293T cell extracts (200 μl per precipitation) or RRL reconstitution reactions (37.5 μl per precipitation) were adjusted to 150 mM NaCl and bound to 10 μl FLAG M2 monoclonal antibody resin (Sigma–Aldrich, St. Louis, MO), 10 μl c-Myc antibody resin (Sigma–Aldrich) or 10 μl streptavidin agarose resin (Sigma–Aldrich) coated with 5′-biotinylated template-antisense oligonucleotide (CTAGACCTGTCATCA GUUAGGGUUAG , where the underlined nucleotides are 2′OMe RNA; [ ]) by end-over-end rotation at room temperature for 2 hr. .. Following binding, the resin was washed three times at room temperature with HLB containing 150 mM NaCl, 0.1% Triton X-100, and 0.2% CHAPS.

    Mass Spectrometry:

    Article Title: Latent Membrane Protein 2A of Epstein-Barr Virus Binds WW Domain E3 Protein-Ubiquitin Ligases That Ubiquitinate B-Cell Tyrosine Kinases
    Article Snippet: Paragraph title: Affinity chromatography and protein sequencing by quadrupole–time-of-flight mass spectrometry. ... For each affinity reagent, a 100-μl-bed volume of streptavidin-agarose resin (Sigma) capable of binding 10 nmol of biotin was washed with 20 mM sodium phosphate buffer (pH 7.0) containing 150 mM NaCl (PBS) and then resuspended in a twofold-molar excess of biotinylated peptide.

    Article Title: The RNA Binding Protein hnRNP Q Modulates the Utilization of Exon 7 in the Survival Motor Neuron 2 (SMN2) Gene ▿) Gene ▿ †
    Article Snippet: After extensive washing with NET-2 buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl) containing 0.25% (wt/vol) NP-40, resin-bound samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, and individual protein bands were analyzed by nanoelectrospray mass spectrometry (MS) (see below). .. To identify HeLa cell proteins that associate with SMN exon 7, 1.2 μg of biotinylated RNA oligonucleotide was conjugated with 20 μl of streptavidin-agarose resin (Sigma).

    Western Blot:

    Article Title: Insights into Brain Glycogen Metabolism
    Article Snippet: Anti-phosphoserine and anti-histidine tag antibodies, l -arabinose, protease inhibitor mixture, nickel-nitrilotriacetic acid Superflow resin, bovine serum albumin, imidazole, streptavidin-agarose resin, phosphoglucomutase, EDTA, glucose-1,6-diphosphate, AMP, ATP, phosphorylase kinase from rabbit muscle, BSA, and glycogen were purchased from Sigma-Aldrich. .. ECL Western blotting detection reagent was purchased from GE Heathcare.

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: After centrifugation, supernatants were incubated with RNA aptamers for 10 min, and then with streptavidin agarose resin (Millipore) for 5 h at 4 °C. .. The binding of USP14 to the 26S proteasome was monitored by western blotting using an antibody against USP14 (Bethyl Laboratories, Inc).

    Article Title: Correlated S-palmitoylation profiling of Snail-induced epithelial to mesenchymal transition
    Article Snippet: Samples were then diluted to 0.5 mg/mL protein and 0.25% SDS in DPBS, and combined with 50 μL of streptavidin-agarose resin (Millipore) rotating for 90 minutes at room temperature. .. Samples were then diluted to 0.5 mg/mL protein and 0.25% SDS in DPBS, and combined with 50 μL of streptavidin-agarose resin (Millipore) rotating for 90 minutes at room temperature.

    RNA Binding Assay:

    Article Title: The RNA Binding Protein hnRNP Q Modulates the Utilization of Exon 7 in the Survival Motor Neuron 2 (SMN2) Gene ▿) Gene ▿ †
    Article Snippet: To identify HeLa cell proteins that associate with SMN exon 7, 1.2 μg of biotinylated RNA oligonucleotide was conjugated with 20 μl of streptavidin-agarose resin (Sigma). .. To compare the SMN RNA binding affinities of hnRNP Q isoforms and fragments, lysates prepared from ∼2 × 106 HEK293 cells containing overexpressed FLAG-tagged protein were subjected to affinity selection as above.

    Protease Inhibitor:

    Article Title: Insights into Brain Glycogen Metabolism
    Article Snippet: .. Anti-phosphoserine and anti-histidine tag antibodies, l -arabinose, protease inhibitor mixture, nickel-nitrilotriacetic acid Superflow resin, bovine serum albumin, imidazole, streptavidin-agarose resin, phosphoglucomutase, EDTA, glucose-1,6-diphosphate, AMP, ATP, phosphorylase kinase from rabbit muscle, BSA, and glycogen were purchased from Sigma-Aldrich. .. Glucose-6-phosphate dehydrogenase was purchased from Roche.

    Buffer Exchange:

    Article Title: Insights into Brain Glycogen Metabolism
    Article Snippet: Biotinylated thrombin was removed from the mixture by the addition of streptavidin-agarose resin (16 μl/unit thrombin, Sigma-Aldrich) for 30 min at 4 °C with agitation. .. The proteins were then subjected to buffer exchange against 20 m m Tris-HCl, pH 6.9, using a PD10 buffer exchange column (GE Healthcare) and concentrated to a concentration of 7 mg/ml.

    Cell Culture:

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: Cells were cultured in 15-cm dishes, harvested in lysis buffer [50 mM NaH2 PO4 pH 7.5, 100 mM NaCl, 10% glycerol, 5 mM MgCl2 , 0.5% NP-40, 5 mM ATP, and 1 mM DTT] containing protease inhibitors, and homogenized using a Dounce homogenizer. .. After centrifugation, supernatants were incubated with streptavidin agarose resin (Millipore) for 5 h at 4 °C.

    Sequencing:

    Article Title: Latent Membrane Protein 2A of Epstein-Barr Virus Binds WW Domain E3 Protein-Ubiquitin Ligases That Ubiquitinate B-Cell Tyrosine Kinases
    Article Snippet: Paragraph title: Affinity chromatography and protein sequencing by quadrupole–time-of-flight mass spectrometry. ... For each affinity reagent, a 100-μl-bed volume of streptavidin-agarose resin (Sigma) capable of binding 10 nmol of biotin was washed with 20 mM sodium phosphate buffer (pH 7.0) containing 150 mM NaCl (PBS) and then resuspended in a twofold-molar excess of biotinylated peptide.

    Sonication:

    Article Title: Nucleotide Excision Repair Is Associated with the Replisome and Its Efficiency Depends on a Direct Interaction between XPA and PCNA
    Article Snippet: Cell pellets were then washed once with 0.5% BSA/PBS and once with PBS, resuspended in lysis buffer (1% SDS, 50 mM Tris (pH 8.0), 1 µg/ml leupeptin, and 1 µg/ml aprotinin) and sonicated. .. Streptavidin–agarose resin (100 µl resin per 2×108 cells, Novagen) was washed twice in lysis buffer and once in PBS.

    Affinity Purification:

    Article Title: Function of glutathione peroxidases in legume root nodules
    Article Snippet: Paragraph title: Affinity purification of biotinylated proteins ... Dry pellets were resuspended in binding buffer consisting of 25mM HEPES (pH 7.7), 1mM EDTA, 100mM NaCl, 0.8% Triton X-100, and 50 μl of streptavidin-agarose resin (Sigma).

    Binding Assay:

    Article Title: Function of glutathione peroxidases in legume root nodules
    Article Snippet: .. Dry pellets were resuspended in binding buffer consisting of 25mM HEPES (pH 7.7), 1mM EDTA, 100mM NaCl, 0.8% Triton X-100, and 50 μl of streptavidin-agarose resin (Sigma). .. Samples were incubated overnight at 4ºC and then the agarose beads were washed ten times with a buffer comprising 25mM HEPES (pH 7.7), 1mM EDTA, 600mM NaCl, and 0.8% Triton X-100.

    Article Title: Latent Membrane Protein 2A of Epstein-Barr Virus Binds WW Domain E3 Protein-Ubiquitin Ligases That Ubiquitinate B-Cell Tyrosine Kinases
    Article Snippet: .. For each affinity reagent, a 100-μl-bed volume of streptavidin-agarose resin (Sigma) capable of binding 10 nmol of biotin was washed with 20 mM sodium phosphate buffer (pH 7.0) containing 150 mM NaCl (PBS) and then resuspended in a twofold-molar excess of biotinylated peptide. .. Following a 15-min incubation and mixing at room temperature, the resin was washed three times with excess PBS.

    Article Title: Single-molecule imaging of telomerase reverse transcriptase in human telomerase holoenzyme and minimal RNP complexes
    Article Snippet: Enrichment of complexes by tagged TERT, tagged TPP1, or hTR template for activity assays HEK 293T cell extracts (200 μl per precipitation) or RRL reconstitution reactions (37.5 μl per precipitation) were adjusted to 150 mM NaCl and bound to 10 μl FLAG M2 monoclonal antibody resin (Sigma–Aldrich, St. Louis, MO), 10 μl c-Myc antibody resin (Sigma–Aldrich) or 10 μl streptavidin agarose resin (Sigma–Aldrich) coated with 5′-biotinylated template-antisense oligonucleotide (CTAGACCTGTCATCA GUUAGGGUUAG , where the underlined nucleotides are 2′OMe RNA; [ ]) by end-over-end rotation at room temperature for 2 hr. .. Following binding, the resin was washed three times at room temperature with HLB containing 150 mM NaCl, 0.1% Triton X-100, and 0.2% CHAPS.

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: Paragraph title: USP14 binding to proteasomes ... After centrifugation, supernatants were incubated with RNA aptamers for 10 min, and then with streptavidin agarose resin (Millipore) for 5 h at 4 °C.

    Article Title: Emergence of a Large-Plaque Variant in Mice Infected with Coxsackievirus B3
    Article Snippet: Paragraph title: Heparin, CAR, and DAF binding assays. ... 35 S-labeled CVB3 (5 × 106 PFU/5,000 cpm) was mixed with heparin agarose resin (Sigma Aldrich) or streptavidin agarose resin (Sigma Aldrich).

    Article Title: Enhanced Functional Potential of Nucleic Acid Aptamer Libraries Patterned to Increase Secondary Structure
    Article Snippet: Streptavidin Selections Round 1: 300 pmol of library mix was denatured in binding buffer (100 mM NaCl, 10 mM MgCl2 , 50 mM Tris, pH 7.8) at 95 °C for 5 min, then cooled on ice for 5 min. .. The DNA was added to 40 μL of streptavidin-agarose resin (Novagen) and incubated with end-over-end rotation for 2 h at 25 °C.

    Nucleic Acid Electrophoresis:

    Article Title: The RNA Binding Protein hnRNP Q Modulates the Utilization of Exon 7 in the Survival Motor Neuron 2 (SMN2) Gene ▿) Gene ▿ †
    Article Snippet: After extensive washing with NET-2 buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl) containing 0.25% (wt/vol) NP-40, resin-bound samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, and individual protein bands were analyzed by nanoelectrospray mass spectrometry (MS) (see below). .. To identify HeLa cell proteins that associate with SMN exon 7, 1.2 μg of biotinylated RNA oligonucleotide was conjugated with 20 μl of streptavidin-agarose resin (Sigma).

    Radioactivity:

    Article Title: Emergence of a Large-Plaque Variant in Mice Infected with Coxsackievirus B3
    Article Snippet: 35 S-labeled CVB3 (5 × 106 PFU/5,000 cpm) was mixed with heparin agarose resin (Sigma Aldrich) or streptavidin agarose resin (Sigma Aldrich). .. Following incubation, resin was pelleted by centrifugation at 2,600 × g for 2 min. Resin was washed three times with PBS+ (PBS supplemented with 100 μg/ml CaCl2 and 100 μg/ml MgCl2 ), and resin-associated radioactivity was quantified by scintillation counting.

    Isolation:

    Article Title: Nucleotide Excision Repair Is Associated with the Replisome and Its Efficiency Depends on a Direct Interaction between XPA and PCNA
    Article Snippet: Paragraph title: Isolation of Proteins on Nascent DNA (iPOND) ... Streptavidin–agarose resin (100 µl resin per 2×108 cells, Novagen) was washed twice in lysis buffer and once in PBS.

    Article Title: Latent Membrane Protein 2A of Epstein-Barr Virus Binds WW Domain E3 Protein-Ubiquitin Ligases That Ubiquitinate B-Cell Tyrosine Kinases
    Article Snippet: For each affinity reagent, a 100-μl-bed volume of streptavidin-agarose resin (Sigma) capable of binding 10 nmol of biotin was washed with 20 mM sodium phosphate buffer (pH 7.0) containing 150 mM NaCl (PBS) and then resuspended in a twofold-molar excess of biotinylated peptide. .. For each affinity isolation, clarified cell lysate from 108 cells was treated with 1.5 nmol (15 μl) of immobilized peptide resin for 90 min at 5°C.

    Purification:

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: Paragraph title: Purification of human proteasomes and preparation of vinylmethylester-proteasomes ... After centrifugation, supernatants were incubated with streptavidin agarose resin (Millipore) for 5 h at 4 °C.

    Article Title: Nucleotide Excision Repair Is Associated with the Replisome and Its Efficiency Depends on a Direct Interaction between XPA and PCNA
    Article Snippet: Samples were centrifuged, filtered through an 80-µm nylon mesh, and diluted 1∶1 with PBS containing 1 µg/ml leupeptin and 1 µg/ml aprotinin prior to purification. .. Streptavidin–agarose resin (100 µl resin per 2×108 cells, Novagen) was washed twice in lysis buffer and once in PBS.

    Article Title: Cryo-EM structure of substrate-bound human telomerase holoenzyme
    Article Snippet: Paragraph title: Telomerase purification ... The prepared whole-cell extract (from 120 plates of adherent cells or 6L of suspension cells) was incubated with streptavidin agarose resin (Sigma-Aldrich) pre-bound to a 5’-biotinylated oligonucleotide at room temperature for 3 hours.

    Selection:

    Article Title: The RNA Binding Protein hnRNP Q Modulates the Utilization of Exon 7 in the Survival Motor Neuron 2 (SMN2) Gene ▿) Gene ▿ †
    Article Snippet: To identify HeLa cell proteins that associate with SMN exon 7, 1.2 μg of biotinylated RNA oligonucleotide was conjugated with 20 μl of streptavidin-agarose resin (Sigma). .. For MS analysis, a 10-fold scale of RNA affinity selection was performed.

    SDS Page:

    Article Title: The RNA Binding Protein hnRNP Q Modulates the Utilization of Exon 7 in the Survival Motor Neuron 2 (SMN2) Gene ▿) Gene ▿ †
    Article Snippet: After extensive washing with NET-2 buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl) containing 0.25% (wt/vol) NP-40, resin-bound samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, and individual protein bands were analyzed by nanoelectrospray mass spectrometry (MS) (see below). .. To identify HeLa cell proteins that associate with SMN exon 7, 1.2 μg of biotinylated RNA oligonucleotide was conjugated with 20 μl of streptavidin-agarose resin (Sigma).

    Article Title: Correlated S-palmitoylation profiling of Snail-induced epithelial to mesenchymal transition
    Article Snippet: Samples were then diluted to 0.5 mg/mL protein and 0.25% SDS in DPBS, and combined with 50 μL of streptavidin-agarose resin (Millipore) rotating for 90 minutes at room temperature. .. Samples were then diluted to 0.5 mg/mL protein and 0.25% SDS in DPBS, and combined with 50 μL of streptavidin-agarose resin (Millipore) rotating for 90 minutes at room temperature.

    Affinity Chromatography:

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: Purification of human proteasomes and preparation of vinylmethylester-proteasomes Human proteasomes were purified by affinity chromatography from a stable HEK293 cell line harboring biotin-tagged human β4 as previously described , with some modifications. .. After centrifugation, supernatants were incubated with streptavidin agarose resin (Millipore) for 5 h at 4 °C.

    Article Title: Latent Membrane Protein 2A of Epstein-Barr Virus Binds WW Domain E3 Protein-Ubiquitin Ligases That Ubiquitinate B-Cell Tyrosine Kinases
    Article Snippet: Paragraph title: Affinity chromatography and protein sequencing by quadrupole–time-of-flight mass spectrometry. ... For each affinity reagent, a 100-μl-bed volume of streptavidin-agarose resin (Sigma) capable of binding 10 nmol of biotin was washed with 20 mM sodium phosphate buffer (pH 7.0) containing 150 mM NaCl (PBS) and then resuspended in a twofold-molar excess of biotinylated peptide.

    Article Title: The RNA Binding Protein hnRNP Q Modulates the Utilization of Exon 7 in the Survival Motor Neuron 2 (SMN2) Gene ▿) Gene ▿ †
    Article Snippet: Paragraph title: RNA affinity chromatography. ... To identify HeLa cell proteins that associate with SMN exon 7, 1.2 μg of biotinylated RNA oligonucleotide was conjugated with 20 μl of streptavidin-agarose resin (Sigma).

    In Vitro:

    Article Title: Ancestral AlaX Editing Enzymes for Control of Genetic Code Fidelity Are Not tRNA-specific *
    Article Snippet: In vitro -transcribed E. coli tRNAPro , P. horikoshii tRNAAla and tRNAThr were 3′ 32 P-labeled using E. coli tRNA nucleotidyltransferase and [α-32 P]ATP (PerkinElmer Life Sciences) as described previously ( ). .. Biotinylated dFx was removed by incubating the reaction mixture with 150 μl of streptavidin-agarose resin (Novagen) for ∼15 min at room temperature, followed by centrifugation in a tabletop centrifuge at 4 °C for 2 min at 2000 × g .

    Concentration Assay:

    Article Title: Nucleotide Excision Repair Is Associated with the Replisome and Its Efficiency Depends on a Direct Interaction between XPA and PCNA
    Article Snippet: Cells were incubated in click reaction buffer for 1–2 h at a concentration of 1×108 cells per 5 ml of click reaction buffer (2 mM CuSO4 , 10 µM biotin-azide, 10 mM Sodium ascorbate). .. Streptavidin–agarose resin (100 µl resin per 2×108 cells, Novagen) was washed twice in lysis buffer and once in PBS.

    Article Title: Distinct tRNA recognition strategies used by a homologous family of editing domains prevent mistranslation
    Article Snippet: Reactions were quenched with 120 μl of 0.3 M NaOAc, pH 5 (0.25 mM final concentration). .. Biotinylated dFx was removed by incubating the reaction mixture with 150 μl of streptavidin agarose resin (Novagen) for ∼15 min at room temperature followed by centrifugation in a table top centrifuge at 4°C for 2 min at 2000×g .

    Article Title: Insights into Brain Glycogen Metabolism
    Article Snippet: Biotinylated thrombin was removed from the mixture by the addition of streptavidin-agarose resin (16 μl/unit thrombin, Sigma-Aldrich) for 30 min at 4 °C with agitation. .. The proteins were then subjected to buffer exchange against 20 m m Tris-HCl, pH 6.9, using a PD10 buffer exchange column (GE Healthcare) and concentrated to a concentration of 7 mg/ml.

    Lysis:

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: Cells were cultured in 15-cm dishes, harvested in lysis buffer [50 mM NaH2 PO4 pH 7.5, 100 mM NaCl, 10% glycerol, 5 mM MgCl2 , 0.5% NP-40, 5 mM ATP, and 1 mM DTT] containing protease inhibitors, and homogenized using a Dounce homogenizer. .. After centrifugation, supernatants were incubated with streptavidin agarose resin (Millipore) for 5 h at 4 °C.

    Article Title: Nucleotide Excision Repair Is Associated with the Replisome and Its Efficiency Depends on a Direct Interaction between XPA and PCNA
    Article Snippet: .. Streptavidin–agarose resin (100 µl resin per 2×108 cells, Novagen) was washed twice in lysis buffer and once in PBS. .. The resin was washed once with lysis buffer, once with 1 M NaCl, and then twice with lysis buffer.

    Article Title: Facilitated Tau Degradation by USP14 Aptamers via Enhanced Proteasome Activity
    Article Snippet: After HEK293 cells were harvested in lysis buffer, cell lysates were homogenized using a 1 mL syringe with a 26G × 1/2′′ needle. .. After centrifugation, supernatants were incubated with RNA aptamers for 10 min, and then with streptavidin agarose resin (Millipore) for 5 h at 4 °C.

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    Millipore streptavidin agarose resin
    Streptavidin Agarose Resin, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore streptavidin sepharose resin
    Streptavidin Sepharose Resin, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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