amylose resin  (New England Biolabs)


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    Name:
    Amylose Resin
    Description:
    Amylose Resin 100 ml
    Catalog Number:
    e8021l
    Price:
    1103
    Size:
    100 ml
    Category:
    Protein Purification Kit Components
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    Structured Review

    New England Biolabs amylose resin
    Amylose Resin
    Amylose Resin 100 ml
    https://www.bioz.com/result/amylose resin/product/New England Biolabs
    Average 90 stars, based on 924 article reviews
    Price from $9.99 to $1999.99
    amylose resin - by Bioz Stars, 2020-01
    90/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Quantitation of SPLUNC1 in saliva with an xMAP particle-based antibody capture and detection immunoassay
    Article Snippet: Briefly, the cDNA for human SPLUNC1 (National Center for Biotechnology Information, NCBI accession number NM 016583) was cloned into the plasmid vector pMAL-c2x (New England Biolabs, Ipswich, MA, USA) for the expression of a fusion protein containing an N-terminal maltose-binding protein tag and a C-terminal 6xHis tag. .. Full-length fusion protein was expressed in the Escherichia coli strain BL21 Star (DE3, Gibco, Invitrogen Corp., Carlsbad, CA, USA). rhSPLUNC1 was purified first, by passing the fusion protein preparation through a column containing amylose resin (New England Biolabs) and then, by passing the fraction containing rhSPLUNC1 through a nickel resin column (Ni Sepharose 6 Fast Flow, GE Healthcare Biosciences Corp., Piscataway, NJ, USA).

    Article Title: Structure and Mechanism of the Lipooligosaccharide Sialyltransferase from Neisseria meningitidis *
    Article Snippet: Paragraph title: Cloning, Expression, and Purification of the MBP-Δ29NST Fusion Protein ... The lysate was centrifuged at 27,000 × g for 30 min, and the resulting supernatant was loaded onto a column (1 column volume of 20 ml) of amylose resin (New England Biolabs) that was equilibrated with the binding buffer (2 m m EDTA (pH 7.5), 20 m m Tris-HCl (pH 7.5), 200 m m NaCl, 10% (v/v) glycerol, and 5 m m β-mercaptoethanol).

    Article Title: An Antibody Directed against the Fusion Peptide of Jun?n Virus Envelope Glycoprotein GPC Inhibits pH-Induced Membrane Fusion ▿
    Article Snippet: The G2 ectodomain region of the molecularly cloned JUNV GPC (amino acids 252 to 424) was appended in frame to the C terminus of maltose-binding protein (MBP) in the pMAL-c2E vector (New England Biolabs) using standard PCR and molecular cloning techniques. .. Aliquots of the cleared lysate were incubated with a 50% slurry of amylose resin (New England BioLabs) for 2 h at 4°C to purify the MBP fusion proteins.

    Expressing:

    Article Title: Dual DNA-binding domains shape the interaction of Brh2 with DNA
    Article Snippet: Brh2NT terminating at residue M551 was purified as a fusion protein with an N-terminal maltose binding protein (MBP)-tag and C-terminal hexahistidine (His)-tag after expression in E. coli . .. Purification involved sequential affinity chromatography on Ni2+ -NTA (nitrilotriacetate) agarose (Qiagen), amylose resin (New England Biolabs), salt gradient elution from monoQ ion exchange resin, and salt gradient elution from HiTrap heparin-agarose using an AKTA FPLC chromatography system (GE Healthcare).

    Article Title: Quantitation of SPLUNC1 in saliva with an xMAP particle-based antibody capture and detection immunoassay
    Article Snippet: Briefly, the cDNA for human SPLUNC1 (National Center for Biotechnology Information, NCBI accession number NM 016583) was cloned into the plasmid vector pMAL-c2x (New England Biolabs, Ipswich, MA, USA) for the expression of a fusion protein containing an N-terminal maltose-binding protein tag and a C-terminal 6xHis tag. .. Full-length fusion protein was expressed in the Escherichia coli strain BL21 Star (DE3, Gibco, Invitrogen Corp., Carlsbad, CA, USA). rhSPLUNC1 was purified first, by passing the fusion protein preparation through a column containing amylose resin (New England Biolabs) and then, by passing the fraction containing rhSPLUNC1 through a nickel resin column (Ni Sepharose 6 Fast Flow, GE Healthcare Biosciences Corp., Piscataway, NJ, USA).

    Article Title: Structure and Mechanism of the Lipooligosaccharide Sialyltransferase from Neisseria meningitidis *
    Article Snippet: Paragraph title: Cloning, Expression, and Purification of the MBP-Δ29NST Fusion Protein ... The lysate was centrifuged at 27,000 × g for 30 min, and the resulting supernatant was loaded onto a column (1 column volume of 20 ml) of amylose resin (New England Biolabs) that was equilibrated with the binding buffer (2 m m EDTA (pH 7.5), 20 m m Tris-HCl (pH 7.5), 200 m m NaCl, 10% (v/v) glycerol, and 5 m m β-mercaptoethanol).

    Article Title: Protein domain definition should allow for conditional disorder
    Article Snippet: Paragraph title: Heterologous protein expression and protein purification ... The MBP fusion proteins were isolated using a bed of amylose resin (New England Biolabs), and eluted in lysis buffer supplemented with 30 m M maltose.

    Article Title: A general path for large-scale solubilization of cellular proteins: From membrane receptors to multiprotein complexes
    Article Snippet: We perform this screen by sonicating 0.5 g of E. coli or Sf9 cells expressing the desired protein in 10 mL of buffer A and aliquot in 6–12 equal fractions. .. We have used the following affinity beads with high success: (1) nickel beads for poly-His tagged proteins (Sigma); (2) antibody beads (IgG, GE healthcare) for protein-A-tagged (PA-tag) proteins; (3) amylose resin (New England Biolabs) for proteins fused to maltose binding protein tag and, (4) chitin beads for proteins purified using the IMPACT-twin purification system (New England Biolabs).

    Article Title: An Antibody Directed against the Fusion Peptide of Jun?n Virus Envelope Glycoprotein GPC Inhibits pH-Induced Membrane Fusion ▿
    Article Snippet: Expression in E. coli TB1 cells (New England Biolabs) was induced by the addition of isopropylthiogalactoside (ITPG), and cells were disrupted using Bugbuster (Pierce) containing 250 units/ml Benzonase (Pierce) and protease inhibitors (leupeptin, pepstatin, aprotinin, and phenylmethylsulfonyl fluoride). .. Aliquots of the cleared lysate were incubated with a 50% slurry of amylose resin (New England BioLabs) for 2 h at 4°C to purify the MBP fusion proteins.

    Article Title: Endoplasmic Reticulum PI(3)P lipid binding targets malaria proteins to the host cell
    Article Snippet: Paragraph title: Expression and purification of recombinant proteins in E. coli ... His-tagged proteins were purified over Pro-Bond resin (Invitrogen), MBP-tagged proteins were purified over Amylose resin (New England Biolabs) and GST-fusions were purified over Glutathione resin (Clontech).

    Ubiquitin Assay:

    Article Title: The Transcription Factor Atf1 Binds and Activates the APC/C Ubiquitin Ligase in Fission Yeast *
    Article Snippet: Paragraph title: Ubiquitylation Assay ... S. pombe MBP-tagged E1 (Uba1) and His-tagged E2s (Ubc1, Ubc4, and Ubc11) were expressed in Escherichia coli and purified using amylose resin (New England Biolab) or Ni-NTA beads (Qiagen) as described by the supplier.

    Incubation:

    Article Title: Resistance protein Pit interacts with the GEF OsSPK1 to activate OsRac1 and trigger rice immunity
    Article Snippet: .. The mixture was applied to 30 µL Amylose resin (New England Biolabs) or Glutathione Sepharose 4B (GE Healthcare) and incubated at 4 °C for 2–3 h. The beads were then washed five times for 5 min each time with pull-down buffer, and the bound proteins were eluted with 100 µl 2× SDS loading buffer and subjected to immunoblot assay with anti-SUMO (A01693; GenScript), anti-GST (sc-138; Santa Cruz Biotechnology), and anti-MBP (E8032S; New England Biolabs) antibodies. .. A. tumefaciens strain GV3101 carrying constructs expressing YFP-OsSPK1 and Pit-GFP mutants were infiltrated into N. benthamiana leaves according to the method described above.

    Article Title: Dual DNA-binding domains shape the interaction of Brh2 with DNA
    Article Snippet: Purification involved sequential affinity chromatography on Ni2+ -NTA (nitrilotriacetate) agarose (Qiagen), amylose resin (New England Biolabs), salt gradient elution from monoQ ion exchange resin, and salt gradient elution from HiTrap heparin-agarose using an AKTA FPLC chromatography system (GE Healthcare). .. Protein complexes were then incubated in buffer A (50mM Tris-HCl, pH 7.5, 200mM KCl, 1 mM DTT, 1mM EDTA, 10% glycerol) containing 20 mM MgCl2 at 37° for 1 hr then loaded onto a column containing amylose resin.

    Article Title: Role of Replication Protein A in Double Holliday Junction Dissolution Mediated by the BLM-Topo III?-RMI1-RMI2 Protein Complex *
    Article Snippet: .. For the affinity pull-down of MBP-tagged proteins, 5 μg of hRPA was incubated with MBP-tagged RMI1425–625 -RMI2, RMI1-RMI2, RMI1Δ301–337 -RMI2, RMI14EA -RMI2, or MBP (5 μg) and mixed with amylose resin (New England Biolabs) to capture the MBP-tagged protein and associated RPA, as above. .. For the affinity pull-down of GST-tagged proteins, 5 μg of hRPA was incubated with 5 μg of GST-tagged RMI1 fragments or 5 μg of GST and mixed with glutathione resin (GE Healthcare) as above.

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin
    Article Snippet: .. Bcl-2-binding assay : A 40 μl volume of 6 μM MBP-Bcl-2 fusion protein (MBP-Bcl-2) was subjected to 40 μl amylose resin (New England Biolabs, Beverly, MA) and incubated for 30 min. .. Thereafter, beads were washed twice with buffer B (25 mM Tris–HCl, pH 7.5, 200 mM NaCl, 1 mM DTT) and subsequently incubated for 1 h with 40 μl reaction mixture containing either FKBP38, FKBP38 (50 μM) and CaM (200 μM), FKBP38 and Ca2+ /CaM, or FKBP38/Ca2+ /CaM and 200 nM GPI1046.

    Article Title: Structure and Mechanism of the Lipooligosaccharide Sialyltransferase from Neisseria meningitidis *
    Article Snippet: The cultures were incubated with shaking (200 rpm at 37 °C) until an A 600 of ∼ 0.2 and were induced by the addition of 1 m m isopropyl β- d -thiogalactopyranoside at 20 °C with further overnight incubation. .. The lysate was centrifuged at 27,000 × g for 30 min, and the resulting supernatant was loaded onto a column (1 column volume of 20 ml) of amylose resin (New England Biolabs) that was equilibrated with the binding buffer (2 m m EDTA (pH 7.5), 20 m m Tris-HCl (pH 7.5), 200 m m NaCl, 10% (v/v) glycerol, and 5 m m β-mercaptoethanol).

    Article Title: A general path for large-scale solubilization of cellular proteins: From membrane receptors to multiprotein complexes
    Article Snippet: Supernatants are mixed with 6–12 aliquots of 50–100 μL of affinity beads (previously equilibrated with buffer A plus 2.5× CMC of the corresponding extracting detergent) and incubated over an hour. .. We have used the following affinity beads with high success: (1) nickel beads for poly-His tagged proteins (Sigma); (2) antibody beads (IgG, GE healthcare) for protein-A-tagged (PA-tag) proteins; (3) amylose resin (New England Biolabs) for proteins fused to maltose binding protein tag and, (4) chitin beads for proteins purified using the IMPACT-twin purification system (New England Biolabs).

    Article Title: An Antibody Directed against the Fusion Peptide of Jun?n Virus Envelope Glycoprotein GPC Inhibits pH-Induced Membrane Fusion ▿
    Article Snippet: .. Aliquots of the cleared lysate were incubated with a 50% slurry of amylose resin (New England BioLabs) for 2 h at 4°C to purify the MBP fusion proteins. ..

    Article Title: Molecular Determinants of Gem Protein Inhibition of P/Q-type Ca2+ Channels *
    Article Snippet: .. MBP fusion peptides were expressed in DE3 and purified by incubation with amylose resin (New England Biolabs). ..

    Stripping Membranes:

    Article Title: Dual DNA-binding domains shape the interaction of Brh2 with DNA
    Article Snippet: Purification involved sequential affinity chromatography on Ni2+ -NTA (nitrilotriacetate) agarose (Qiagen), amylose resin (New England Biolabs), salt gradient elution from monoQ ion exchange resin, and salt gradient elution from HiTrap heparin-agarose using an AKTA FPLC chromatography system (GE Healthcare). .. Brh2 and Brh2CT proteins tagged with MBP were purified after obtaining the respective heterodimeric forms in complex with His-tagged Dss1 as described previously [ , ], then stripping off and discarding the His-Dss1.

    Mass Spectrometry:

    Article Title: Protein domain definition should allow for conditional disorder
    Article Snippet: The MBP fusion proteins were isolated using a bed of amylose resin (New England Biolabs), and eluted in lysis buffer supplemented with 30 m M maltose. .. Masses of all foot domains were confirmed using MALDI-TOF mass spectrometry.

    Western Blot:

    Article Title: Stepwise Activity of ClpY (HslU) Mutants in the Processive Degradation of Escherichia coli ClpYQ (HslUV) Protease Substrates ▿
    Article Snippet: ClpY and its above derivative mutants were each purified using His tag affinity purification methods, whereas MBP-SulA was purified using amylose resin (NEB) methods and the concentration of each protein was determined (see Materials and Methods). .. As shown in , each of the various ClpY mutants was capable of pulling down with MBP-SulA, seen in the SDS-PAGE gel and in the Western blot analyses.

    Article Title: A general path for large-scale solubilization of cellular proteins: From membrane receptors to multiprotein complexes
    Article Snippet: We have used the following affinity beads with high success: (1) nickel beads for poly-His tagged proteins (Sigma); (2) antibody beads (IgG, GE healthcare) for protein-A-tagged (PA-tag) proteins; (3) amylose resin (New England Biolabs) for proteins fused to maltose binding protein tag and, (4) chitin beads for proteins purified using the IMPACT-twin purification system (New England Biolabs). .. Alternatively, for protein expressing at low levels, we perform western blots to quantify purification efficiency.

    Article Title: An Antibody Directed against the Fusion Peptide of Jun?n Virus Envelope Glycoprotein GPC Inhibits pH-Induced Membrane Fusion ▿
    Article Snippet: Aliquots of the cleared lysate were incubated with a 50% slurry of amylose resin (New England BioLabs) for 2 h at 4°C to purify the MBP fusion proteins. .. Proteins were resolved using 4-to-12% NuPAGE bis-Tris gels (Invitrogen) and detected by SYPRO Red protein stain (Molecular Probes) or by Western blot analysis.

    Recombinase Polymerase Amplification:

    Article Title: Role of Replication Protein A in Double Holliday Junction Dissolution Mediated by the BLM-Topo III?-RMI1-RMI2 Protein Complex *
    Article Snippet: .. For the affinity pull-down of MBP-tagged proteins, 5 μg of hRPA was incubated with MBP-tagged RMI1425–625 -RMI2, RMI1-RMI2, RMI1Δ301–337 -RMI2, RMI14EA -RMI2, or MBP (5 μg) and mixed with amylose resin (New England Biolabs) to capture the MBP-tagged protein and associated RPA, as above. .. For the affinity pull-down of GST-tagged proteins, 5 μg of hRPA was incubated with 5 μg of GST-tagged RMI1 fragments or 5 μg of GST and mixed with glutathione resin (GE Healthcare) as above.

    Flow Cytometry:

    Article Title: Quantitation of SPLUNC1 in saliva with an xMAP particle-based antibody capture and detection immunoassay
    Article Snippet: .. Full-length fusion protein was expressed in the Escherichia coli strain BL21 Star (DE3, Gibco, Invitrogen Corp., Carlsbad, CA, USA). rhSPLUNC1 was purified first, by passing the fusion protein preparation through a column containing amylose resin (New England Biolabs) and then, by passing the fraction containing rhSPLUNC1 through a nickel resin column (Ni Sepharose 6 Fast Flow, GE Healthcare Biosciences Corp., Piscataway, NJ, USA). .. The maltose-binding protein tag was cleaved from the purified rhSPLUNC1 using Factor Xa protease (New England Biolabs).

    Article Title: Structural Basis for Influenza Virus NS1 Protein Block of mRNA Nuclear Export
    Article Snippet: .. 50xAdvantage Polymerase mix (Clontech (EMD), 639202); dNTP’s (Clontech (EMD), 639125); BamHI-HF (New England Biolabs (NEB), R3136T); NotI (New England Biolabs (NEB), R0189S); SmaI (New England Biolabs (NEB), R0141); XhoI (New England Biolabs (NEB), R0146S); T4 DNA Ligase (New England Biolabs (NEB), M0202); QIAquick Gel Extraction Kit (Qiagen, 28704); NEB® 5-alpha Competent E. coli (New England Biolabs (NEB), ); Rosetta (DE3) Competent Cells (EMD Millipore, 70954); SOC Outgrowth Medium (New England Biolabs (NEB), B9020); QIAprep Spin Miniprep Kit (250) (Qiagen, 27106); Lipofectamine® RNAiMAX Transfection Reagent (Thermo Fisher Scientific, 13778150); RNeasy Plus Mini Kit (Qiagen, 74134); Random Hexamers (50 μM) (Thermo Fisher Scientific, N8080127); Protector RNase Inhibitor (Roche, 03335402001); SuperScript II Reverse Transcriptase (Thermo Fisher Scientific, 18064014); LIGHTCYCLER 480 SYBR GREEN I MASTER (Roche, 04707516001); LightCycler® 480 Multiwell Plate 96, White (Roche, 04729692001); NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, 78833); Complete EDTA-free protease inhibitor tablets (Sigma-Aldrich, 11873580001); TransIT-X2® Dynamic Delivery System (Mirus Bio, MIR6000); SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific, 34096); L-Glutathione reduced (Sigma-Aldrich, G4251–25G); Ampicillin (Sigma-Aldrich, A-9518); Kanamycin Monosulfate (Gold Biotechnology, K-120–10); IPTG (Gold Biotechnology, I2481C50); Imidazole (Sigma-Aldrich, 56750); PMSF (RPI, ); Aprotinin (Santa Cruz Biotechnology, sc-3595); Leupeptin (Santa Cruz Biotechnology, sc295358); Pepstatin A (Thermo Fisher Scientific, ); Glutathione Sepharose 4B (GE Healthcare, 17–0756-01); Amylose Resin (New England Biolabs (NEB), E8021S); Ni-NTA Agarose (Qiagen, 30210); Mono Q 5/50 GL (GE Healthcare, 17-5166-01); HiTrap SP HP (GE Healthcare, 17-1151-01); Superdex 200 HR 10/30 (GE Healthcare, 17-1088-01); EasyTag EXPRESS35 S Protein Labeling Mix, [35S]-, 7mCi (PerkinElmer, NEG772007MC); T7 RiboMAX™ Express Large Scale RNA Production System (Promega, P1320); Protein G Sepharose® 4 Fast Flow (GE Healthcare, 17-0618-01); Hoechst 33258 (Molecular Probes/Life Technologies); M mRNA probes (Biosearch Technologies) ; ProLong Gold antifade reagent (Life Technologies, ). ..

    Chromatography:

    Article Title: Dual DNA-binding domains shape the interaction of Brh2 with DNA
    Article Snippet: .. Purification involved sequential affinity chromatography on Ni2+ -NTA (nitrilotriacetate) agarose (Qiagen), amylose resin (New England Biolabs), salt gradient elution from monoQ ion exchange resin, and salt gradient elution from HiTrap heparin-agarose using an AKTA FPLC chromatography system (GE Healthcare). .. Brh2 and Brh2CT proteins tagged with MBP were purified after obtaining the respective heterodimeric forms in complex with His-tagged Dss1 as described previously [ , ], then stripping off and discarding the His-Dss1.

    Article Title: Protein domain definition should allow for conditional disorder
    Article Snippet: The MBP fusion proteins were isolated using a bed of amylose resin (New England Biolabs), and eluted in lysis buffer supplemented with 30 m M maltose. .. Proteins were further purified using anion exchange chromatography, binding them to Q Sepharose HP resin (Amersham Biosciences) buffered with 12.5 m M Tris/HCl pH 8.5, 50 m M NaCl, and then displacing them with a linear salt gradient.

    Protease Inhibitor:

    Article Title: Structure and Mechanism of the Lipooligosaccharide Sialyltransferase from Neisseria meningitidis *
    Article Snippet: The cell pellet was resuspended in lysis buffer (2 m m EDTA (pH 7.5), 20 m m Tris-HCl (pH 7.5), 200 m m NaCl, 10% (v/v) glycerol) supplemented with a Roche Applied Science EDTA-free protease inhibitor mixture tablet and lysed at 20,000 p.s.i. using the Avestin Emulsiflex homogenizer. .. The lysate was centrifuged at 27,000 × g for 30 min, and the resulting supernatant was loaded onto a column (1 column volume of 20 ml) of amylose resin (New England Biolabs) that was equilibrated with the binding buffer (2 m m EDTA (pH 7.5), 20 m m Tris-HCl (pH 7.5), 200 m m NaCl, 10% (v/v) glycerol, and 5 m m β-mercaptoethanol).

    Article Title: Structural Basis for Influenza Virus NS1 Protein Block of mRNA Nuclear Export
    Article Snippet: .. 50xAdvantage Polymerase mix (Clontech (EMD), 639202); dNTP’s (Clontech (EMD), 639125); BamHI-HF (New England Biolabs (NEB), R3136T); NotI (New England Biolabs (NEB), R0189S); SmaI (New England Biolabs (NEB), R0141); XhoI (New England Biolabs (NEB), R0146S); T4 DNA Ligase (New England Biolabs (NEB), M0202); QIAquick Gel Extraction Kit (Qiagen, 28704); NEB® 5-alpha Competent E. coli (New England Biolabs (NEB), ); Rosetta (DE3) Competent Cells (EMD Millipore, 70954); SOC Outgrowth Medium (New England Biolabs (NEB), B9020); QIAprep Spin Miniprep Kit (250) (Qiagen, 27106); Lipofectamine® RNAiMAX Transfection Reagent (Thermo Fisher Scientific, 13778150); RNeasy Plus Mini Kit (Qiagen, 74134); Random Hexamers (50 μM) (Thermo Fisher Scientific, N8080127); Protector RNase Inhibitor (Roche, 03335402001); SuperScript II Reverse Transcriptase (Thermo Fisher Scientific, 18064014); LIGHTCYCLER 480 SYBR GREEN I MASTER (Roche, 04707516001); LightCycler® 480 Multiwell Plate 96, White (Roche, 04729692001); NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, 78833); Complete EDTA-free protease inhibitor tablets (Sigma-Aldrich, 11873580001); TransIT-X2® Dynamic Delivery System (Mirus Bio, MIR6000); SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific, 34096); L-Glutathione reduced (Sigma-Aldrich, G4251–25G); Ampicillin (Sigma-Aldrich, A-9518); Kanamycin Monosulfate (Gold Biotechnology, K-120–10); IPTG (Gold Biotechnology, I2481C50); Imidazole (Sigma-Aldrich, 56750); PMSF (RPI, ); Aprotinin (Santa Cruz Biotechnology, sc-3595); Leupeptin (Santa Cruz Biotechnology, sc295358); Pepstatin A (Thermo Fisher Scientific, ); Glutathione Sepharose 4B (GE Healthcare, 17–0756-01); Amylose Resin (New England Biolabs (NEB), E8021S); Ni-NTA Agarose (Qiagen, 30210); Mono Q 5/50 GL (GE Healthcare, 17-5166-01); HiTrap SP HP (GE Healthcare, 17-1151-01); Superdex 200 HR 10/30 (GE Healthcare, 17-1088-01); EasyTag EXPRESS35 S Protein Labeling Mix, [35S]-, 7mCi (PerkinElmer, NEG772007MC); T7 RiboMAX™ Express Large Scale RNA Production System (Promega, P1320); Protein G Sepharose® 4 Fast Flow (GE Healthcare, 17-0618-01); Hoechst 33258 (Molecular Probes/Life Technologies); M mRNA probes (Biosearch Technologies) ; ProLong Gold antifade reagent (Life Technologies, ). ..

    Dissection:

    Article Title: An Antibody Directed against the Fusion Peptide of Jun?n Virus Envelope Glycoprotein GPC Inhibits pH-Induced Membrane Fusion ▿
    Article Snippet: Paragraph title: Dissection of MAb epitopes. ... Aliquots of the cleared lysate were incubated with a 50% slurry of amylose resin (New England BioLabs) for 2 h at 4°C to purify the MBP fusion proteins.

    Polymerase Chain Reaction:

    Article Title: An Antibody Directed against the Fusion Peptide of Jun?n Virus Envelope Glycoprotein GPC Inhibits pH-Induced Membrane Fusion ▿
    Article Snippet: The G2 ectodomain region of the molecularly cloned JUNV GPC (amino acids 252 to 424) was appended in frame to the C terminus of maltose-binding protein (MBP) in the pMAL-c2E vector (New England Biolabs) using standard PCR and molecular cloning techniques. .. Aliquots of the cleared lysate were incubated with a 50% slurry of amylose resin (New England BioLabs) for 2 h at 4°C to purify the MBP fusion proteins.

    Sonication:

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin
    Article Snippet: Bcl-2-binding assay : A 40 μl volume of 6 μM MBP-Bcl-2 fusion protein (MBP-Bcl-2) was subjected to 40 μl amylose resin (New England Biolabs, Beverly, MA) and incubated for 30 min. .. To prepare SH-SY5Y crude cell extract, cells were harvested, centrifuged for 10 min at 2000 g and cell pellets were resuspended in the same volume of hypotonic lysis buffer, followed by sonication.

    Affinity Purification:

    Article Title: The Transcription Factor Atf1 Binds and Activates the APC/C Ubiquitin Ligase in Fission Yeast *
    Article Snippet: S. pombe MBP-tagged E1 (Uba1) and His-tagged E2s (Ubc1, Ubc4, and Ubc11) were expressed in Escherichia coli and purified using amylose resin (New England Biolab) or Ni-NTA beads (Qiagen) as described by the supplier. .. S. pombe APC/C was purified from apc4 / lid1 + -TAP strain using tandem affinity purification ( ).

    Article Title: Stepwise Activity of ClpY (HslU) Mutants in the Processive Degradation of Escherichia coli ClpYQ (HslUV) Protease Substrates ▿
    Article Snippet: .. ClpY and its above derivative mutants were each purified using His tag affinity purification methods, whereas MBP-SulA was purified using amylose resin (NEB) methods and the concentration of each protein was determined (see Materials and Methods). .. Each ClpY mutant protein and the wild-type ClpY were equivalently tested for their own pulling down with MBP-SulA in the presence of ATP.

    Recombinant:

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin
    Article Snippet: Subsequently, 30 μg recombinant FKBP38 was incubated with CaM-Sepharose. .. Bcl-2-binding assay : A 40 μl volume of 6 μM MBP-Bcl-2 fusion protein (MBP-Bcl-2) was subjected to 40 μl amylose resin (New England Biolabs, Beverly, MA) and incubated for 30 min.

    Article Title: Quantitation of SPLUNC1 in saliva with an xMAP particle-based antibody capture and detection immunoassay
    Article Snippet: Recombinant human SPLUNC1 (rhSPLUNC1) was prepared as previously described . .. Full-length fusion protein was expressed in the Escherichia coli strain BL21 Star (DE3, Gibco, Invitrogen Corp., Carlsbad, CA, USA). rhSPLUNC1 was purified first, by passing the fusion protein preparation through a column containing amylose resin (New England Biolabs) and then, by passing the fraction containing rhSPLUNC1 through a nickel resin column (Ni Sepharose 6 Fast Flow, GE Healthcare Biosciences Corp., Piscataway, NJ, USA).

    Article Title: Endoplasmic Reticulum PI(3)P lipid binding targets malaria proteins to the host cell
    Article Snippet: Paragraph title: Expression and purification of recombinant proteins in E. coli ... His-tagged proteins were purified over Pro-Bond resin (Invitrogen), MBP-tagged proteins were purified over Amylose resin (New England Biolabs) and GST-fusions were purified over Glutathione resin (Clontech).

    Molecular Cloning:

    Article Title: An Antibody Directed against the Fusion Peptide of Jun?n Virus Envelope Glycoprotein GPC Inhibits pH-Induced Membrane Fusion ▿
    Article Snippet: The G2 ectodomain region of the molecularly cloned JUNV GPC (amino acids 252 to 424) was appended in frame to the C terminus of maltose-binding protein (MBP) in the pMAL-c2E vector (New England Biolabs) using standard PCR and molecular cloning techniques. .. Aliquots of the cleared lysate were incubated with a 50% slurry of amylose resin (New England BioLabs) for 2 h at 4°C to purify the MBP fusion proteins.

    Mutagenesis:

    Article Title: Stepwise Activity of ClpY (HslU) Mutants in the Processive Degradation of Escherichia coli ClpYQ (HslUV) Protease Substrates ▿
    Article Snippet: ClpY and its above derivative mutants were each purified using His tag affinity purification methods, whereas MBP-SulA was purified using amylose resin (NEB) methods and the concentration of each protein was determined (see Materials and Methods). .. Each ClpY mutant protein and the wild-type ClpY were equivalently tested for their own pulling down with MBP-SulA in the presence of ATP.

    Isolation:

    Article Title: Protein domain definition should allow for conditional disorder
    Article Snippet: .. The MBP fusion proteins were isolated using a bed of amylose resin (New England Biolabs), and eluted in lysis buffer supplemented with 30 m M maltose. .. Proteins were further purified using anion exchange chromatography, binding them to Q Sepharose HP resin (Amersham Biosciences) buffered with 12.5 m M Tris/HCl pH 8.5, 50 m M NaCl, and then displacing them with a linear salt gradient.

    Transfection:

    Article Title: Structural Basis for Influenza Virus NS1 Protein Block of mRNA Nuclear Export
    Article Snippet: .. 50xAdvantage Polymerase mix (Clontech (EMD), 639202); dNTP’s (Clontech (EMD), 639125); BamHI-HF (New England Biolabs (NEB), R3136T); NotI (New England Biolabs (NEB), R0189S); SmaI (New England Biolabs (NEB), R0141); XhoI (New England Biolabs (NEB), R0146S); T4 DNA Ligase (New England Biolabs (NEB), M0202); QIAquick Gel Extraction Kit (Qiagen, 28704); NEB® 5-alpha Competent E. coli (New England Biolabs (NEB), ); Rosetta (DE3) Competent Cells (EMD Millipore, 70954); SOC Outgrowth Medium (New England Biolabs (NEB), B9020); QIAprep Spin Miniprep Kit (250) (Qiagen, 27106); Lipofectamine® RNAiMAX Transfection Reagent (Thermo Fisher Scientific, 13778150); RNeasy Plus Mini Kit (Qiagen, 74134); Random Hexamers (50 μM) (Thermo Fisher Scientific, N8080127); Protector RNase Inhibitor (Roche, 03335402001); SuperScript II Reverse Transcriptase (Thermo Fisher Scientific, 18064014); LIGHTCYCLER 480 SYBR GREEN I MASTER (Roche, 04707516001); LightCycler® 480 Multiwell Plate 96, White (Roche, 04729692001); NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, 78833); Complete EDTA-free protease inhibitor tablets (Sigma-Aldrich, 11873580001); TransIT-X2® Dynamic Delivery System (Mirus Bio, MIR6000); SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific, 34096); L-Glutathione reduced (Sigma-Aldrich, G4251–25G); Ampicillin (Sigma-Aldrich, A-9518); Kanamycin Monosulfate (Gold Biotechnology, K-120–10); IPTG (Gold Biotechnology, I2481C50); Imidazole (Sigma-Aldrich, 56750); PMSF (RPI, ); Aprotinin (Santa Cruz Biotechnology, sc-3595); Leupeptin (Santa Cruz Biotechnology, sc295358); Pepstatin A (Thermo Fisher Scientific, ); Glutathione Sepharose 4B (GE Healthcare, 17–0756-01); Amylose Resin (New England Biolabs (NEB), E8021S); Ni-NTA Agarose (Qiagen, 30210); Mono Q 5/50 GL (GE Healthcare, 17-5166-01); HiTrap SP HP (GE Healthcare, 17-1151-01); Superdex 200 HR 10/30 (GE Healthcare, 17-1088-01); EasyTag EXPRESS35 S Protein Labeling Mix, [35S]-, 7mCi (PerkinElmer, NEG772007MC); T7 RiboMAX™ Express Large Scale RNA Production System (Promega, P1320); Protein G Sepharose® 4 Fast Flow (GE Healthcare, 17-0618-01); Hoechst 33258 (Molecular Probes/Life Technologies); M mRNA probes (Biosearch Technologies) ; ProLong Gold antifade reagent (Life Technologies, ). ..

    Labeling:

    Article Title: Structural Basis for Influenza Virus NS1 Protein Block of mRNA Nuclear Export
    Article Snippet: .. 50xAdvantage Polymerase mix (Clontech (EMD), 639202); dNTP’s (Clontech (EMD), 639125); BamHI-HF (New England Biolabs (NEB), R3136T); NotI (New England Biolabs (NEB), R0189S); SmaI (New England Biolabs (NEB), R0141); XhoI (New England Biolabs (NEB), R0146S); T4 DNA Ligase (New England Biolabs (NEB), M0202); QIAquick Gel Extraction Kit (Qiagen, 28704); NEB® 5-alpha Competent E. coli (New England Biolabs (NEB), ); Rosetta (DE3) Competent Cells (EMD Millipore, 70954); SOC Outgrowth Medium (New England Biolabs (NEB), B9020); QIAprep Spin Miniprep Kit (250) (Qiagen, 27106); Lipofectamine® RNAiMAX Transfection Reagent (Thermo Fisher Scientific, 13778150); RNeasy Plus Mini Kit (Qiagen, 74134); Random Hexamers (50 μM) (Thermo Fisher Scientific, N8080127); Protector RNase Inhibitor (Roche, 03335402001); SuperScript II Reverse Transcriptase (Thermo Fisher Scientific, 18064014); LIGHTCYCLER 480 SYBR GREEN I MASTER (Roche, 04707516001); LightCycler® 480 Multiwell Plate 96, White (Roche, 04729692001); NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, 78833); Complete EDTA-free protease inhibitor tablets (Sigma-Aldrich, 11873580001); TransIT-X2® Dynamic Delivery System (Mirus Bio, MIR6000); SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific, 34096); L-Glutathione reduced (Sigma-Aldrich, G4251–25G); Ampicillin (Sigma-Aldrich, A-9518); Kanamycin Monosulfate (Gold Biotechnology, K-120–10); IPTG (Gold Biotechnology, I2481C50); Imidazole (Sigma-Aldrich, 56750); PMSF (RPI, ); Aprotinin (Santa Cruz Biotechnology, sc-3595); Leupeptin (Santa Cruz Biotechnology, sc295358); Pepstatin A (Thermo Fisher Scientific, ); Glutathione Sepharose 4B (GE Healthcare, 17–0756-01); Amylose Resin (New England Biolabs (NEB), E8021S); Ni-NTA Agarose (Qiagen, 30210); Mono Q 5/50 GL (GE Healthcare, 17-5166-01); HiTrap SP HP (GE Healthcare, 17-1151-01); Superdex 200 HR 10/30 (GE Healthcare, 17-1088-01); EasyTag EXPRESS35 S Protein Labeling Mix, [35S]-, 7mCi (PerkinElmer, NEG772007MC); T7 RiboMAX™ Express Large Scale RNA Production System (Promega, P1320); Protein G Sepharose® 4 Fast Flow (GE Healthcare, 17-0618-01); Hoechst 33258 (Molecular Probes/Life Technologies); M mRNA probes (Biosearch Technologies) ; ProLong Gold antifade reagent (Life Technologies, ). ..

    Purification:

    Article Title: Dual DNA-binding domains shape the interaction of Brh2 with DNA
    Article Snippet: .. Purification involved sequential affinity chromatography on Ni2+ -NTA (nitrilotriacetate) agarose (Qiagen), amylose resin (New England Biolabs), salt gradient elution from monoQ ion exchange resin, and salt gradient elution from HiTrap heparin-agarose using an AKTA FPLC chromatography system (GE Healthcare). .. Brh2 and Brh2CT proteins tagged with MBP were purified after obtaining the respective heterodimeric forms in complex with His-tagged Dss1 as described previously [ , ], then stripping off and discarding the His-Dss1.

    Article Title: The Transcription Factor Atf1 Binds and Activates the APC/C Ubiquitin Ligase in Fission Yeast *
    Article Snippet: .. S. pombe MBP-tagged E1 (Uba1) and His-tagged E2s (Ubc1, Ubc4, and Ubc11) were expressed in Escherichia coli and purified using amylose resin (New England Biolab) or Ni-NTA beads (Qiagen) as described by the supplier. .. S. pombe APC/C was purified from apc4 / lid1 + -TAP strain using tandem affinity purification ( ).

    Article Title: Quantitation of SPLUNC1 in saliva with an xMAP particle-based antibody capture and detection immunoassay
    Article Snippet: .. Full-length fusion protein was expressed in the Escherichia coli strain BL21 Star (DE3, Gibco, Invitrogen Corp., Carlsbad, CA, USA). rhSPLUNC1 was purified first, by passing the fusion protein preparation through a column containing amylose resin (New England Biolabs) and then, by passing the fraction containing rhSPLUNC1 through a nickel resin column (Ni Sepharose 6 Fast Flow, GE Healthcare Biosciences Corp., Piscataway, NJ, USA). .. The maltose-binding protein tag was cleaved from the purified rhSPLUNC1 using Factor Xa protease (New England Biolabs).

    Article Title: Structure and Mechanism of the Lipooligosaccharide Sialyltransferase from Neisseria meningitidis *
    Article Snippet: Paragraph title: Cloning, Expression, and Purification of the MBP-Δ29NST Fusion Protein ... The lysate was centrifuged at 27,000 × g for 30 min, and the resulting supernatant was loaded onto a column (1 column volume of 20 ml) of amylose resin (New England Biolabs) that was equilibrated with the binding buffer (2 m m EDTA (pH 7.5), 20 m m Tris-HCl (pH 7.5), 200 m m NaCl, 10% (v/v) glycerol, and 5 m m β-mercaptoethanol).

    Article Title: Protein domain definition should allow for conditional disorder
    Article Snippet: The Mapuera virus foot domain was purified using an alternative procedure, due to its propensity to bind irreversibly to chromatographic resins. .. The MBP fusion proteins were isolated using a bed of amylose resin (New England Biolabs), and eluted in lysis buffer supplemented with 30 m M maltose.

    Article Title: Stepwise Activity of ClpY (HslU) Mutants in the Processive Degradation of Escherichia coli ClpYQ (HslUV) Protease Substrates ▿
    Article Snippet: .. ClpY and its above derivative mutants were each purified using His tag affinity purification methods, whereas MBP-SulA was purified using amylose resin (NEB) methods and the concentration of each protein was determined (see Materials and Methods). .. Each ClpY mutant protein and the wild-type ClpY were equivalently tested for their own pulling down with MBP-SulA in the presence of ATP.

    Article Title: A general path for large-scale solubilization of cellular proteins: From membrane receptors to multiprotein complexes
    Article Snippet: .. We have used the following affinity beads with high success: (1) nickel beads for poly-His tagged proteins (Sigma); (2) antibody beads (IgG, GE healthcare) for protein-A-tagged (PA-tag) proteins; (3) amylose resin (New England Biolabs) for proteins fused to maltose binding protein tag and, (4) chitin beads for proteins purified using the IMPACT-twin purification system (New England Biolabs). .. Following incubation, beads are thoroughly washed several times with buffer A plus 2.5× CMC of the corresponding detergent, and eluted (or boiled); individual fractions are subject to SDS–PAGE to quantify protein extraction and quality of purification.

    Article Title: Endoplasmic Reticulum PI(3)P lipid binding targets malaria proteins to the host cell
    Article Snippet: .. His-tagged proteins were purified over Pro-Bond resin (Invitrogen), MBP-tagged proteins were purified over Amylose resin (New England Biolabs) and GST-fusions were purified over Glutathione resin (Clontech). ..

    Article Title: Molecular Determinants of Gem Protein Inhibition of P/Q-type Ca2+ Channels *
    Article Snippet: .. MBP fusion peptides were expressed in DE3 and purified by incubation with amylose resin (New England Biolabs). ..

    Article Title: Control of Heme Homeostasis in Corynebacterium glutamicum by the Two-Component System HrrSA ▿ by the Two-Component System HrrSA ▿ †
    Article Snippet: .. MBP-HrrSΔ1-248 present in the supernatant after ultracentrifugation was purified by affinity chromatography on amylose resin (New England BioLabs) equilibrated with TNM buffer (50 mM Tris-HCl, pH 8.0, 200 mM NaCl, 5 mM MgCl2 ). ..

    Protein Purification:

    Article Title: Dual DNA-binding domains shape the interaction of Brh2 with DNA
    Article Snippet: Paragraph title: 2.1 Protein purification and manipulation ... Purification involved sequential affinity chromatography on Ni2+ -NTA (nitrilotriacetate) agarose (Qiagen), amylose resin (New England Biolabs), salt gradient elution from monoQ ion exchange resin, and salt gradient elution from HiTrap heparin-agarose using an AKTA FPLC chromatography system (GE Healthcare).

    Article Title: Protein domain definition should allow for conditional disorder
    Article Snippet: Paragraph title: Heterologous protein expression and protein purification ... The MBP fusion proteins were isolated using a bed of amylose resin (New England Biolabs), and eluted in lysis buffer supplemented with 30 m M maltose.

    Protein Extraction:

    Article Title: A general path for large-scale solubilization of cellular proteins: From membrane receptors to multiprotein complexes
    Article Snippet: We have used the following affinity beads with high success: (1) nickel beads for poly-His tagged proteins (Sigma); (2) antibody beads (IgG, GE healthcare) for protein-A-tagged (PA-tag) proteins; (3) amylose resin (New England Biolabs) for proteins fused to maltose binding protein tag and, (4) chitin beads for proteins purified using the IMPACT-twin purification system (New England Biolabs). .. Following incubation, beads are thoroughly washed several times with buffer A plus 2.5× CMC of the corresponding detergent, and eluted (or boiled); individual fractions are subject to SDS–PAGE to quantify protein extraction and quality of purification.

    Affinity Chromatography:

    Article Title: Dual DNA-binding domains shape the interaction of Brh2 with DNA
    Article Snippet: .. Purification involved sequential affinity chromatography on Ni2+ -NTA (nitrilotriacetate) agarose (Qiagen), amylose resin (New England Biolabs), salt gradient elution from monoQ ion exchange resin, and salt gradient elution from HiTrap heparin-agarose using an AKTA FPLC chromatography system (GE Healthcare). .. Brh2 and Brh2CT proteins tagged with MBP were purified after obtaining the respective heterodimeric forms in complex with His-tagged Dss1 as described previously [ , ], then stripping off and discarding the His-Dss1.

    Article Title: Control of Heme Homeostasis in Corynebacterium glutamicum by the Two-Component System HrrSA ▿ by the Two-Component System HrrSA ▿ †
    Article Snippet: .. MBP-HrrSΔ1-248 present in the supernatant after ultracentrifugation was purified by affinity chromatography on amylose resin (New England BioLabs) equilibrated with TNM buffer (50 mM Tris-HCl, pH 8.0, 200 mM NaCl, 5 mM MgCl2 ). ..

    Fast Protein Liquid Chromatography:

    Article Title: Dual DNA-binding domains shape the interaction of Brh2 with DNA
    Article Snippet: .. Purification involved sequential affinity chromatography on Ni2+ -NTA (nitrilotriacetate) agarose (Qiagen), amylose resin (New England Biolabs), salt gradient elution from monoQ ion exchange resin, and salt gradient elution from HiTrap heparin-agarose using an AKTA FPLC chromatography system (GE Healthcare). .. Brh2 and Brh2CT proteins tagged with MBP were purified after obtaining the respective heterodimeric forms in complex with His-tagged Dss1 as described previously [ , ], then stripping off and discarding the His-Dss1.

    Staining:

    Article Title: Role of Replication Protein A in Double Holliday Junction Dissolution Mediated by the BLM-Topo III?-RMI1-RMI2 Protein Complex *
    Article Snippet: The supernatant, wash, and SDS eluate, 10 μl of each, were analyzed by 7.5% SDS-PAGE and Coomassie Blue staining. .. For the affinity pull-down of MBP-tagged proteins, 5 μg of hRPA was incubated with MBP-tagged RMI1425–625 -RMI2, RMI1-RMI2, RMI1Δ301–337 -RMI2, RMI14EA -RMI2, or MBP (5 μg) and mixed with amylose resin (New England Biolabs) to capture the MBP-tagged protein and associated RPA, as above.

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin
    Article Snippet: Sepharose was washed and bound proteins were analyzed by 12.5% SDS–PAGE and Coomassie blue staining. .. Bcl-2-binding assay : A 40 μl volume of 6 μM MBP-Bcl-2 fusion protein (MBP-Bcl-2) was subjected to 40 μl amylose resin (New England Biolabs, Beverly, MA) and incubated for 30 min.

    Article Title: Protein domain definition should allow for conditional disorder
    Article Snippet: The MBP fusion proteins were isolated using a bed of amylose resin (New England Biolabs), and eluted in lysis buffer supplemented with 30 m M maltose. .. Protein purity was assessed using SDS-PAGE, coupled with colloidal Coomassie staining for protein detection.

    Article Title: An Antibody Directed against the Fusion Peptide of Jun?n Virus Envelope Glycoprotein GPC Inhibits pH-Induced Membrane Fusion ▿
    Article Snippet: Aliquots of the cleared lysate were incubated with a 50% slurry of amylose resin (New England BioLabs) for 2 h at 4°C to purify the MBP fusion proteins. .. Proteins were resolved using 4-to-12% NuPAGE bis-Tris gels (Invitrogen) and detected by SYPRO Red protein stain (Molecular Probes) or by Western blot analysis.

    Article Title: Molecular Determinants of Gem Protein Inhibition of P/Q-type Ca2+ Channels *
    Article Snippet: MBP fusion peptides were expressed in DE3 and purified by incubation with amylose resin (New England Biolabs). .. The elution was detected with Coomassie Blue staining on SDS-PAGE gel.

    SDS Page:

    Article Title: Role of Replication Protein A in Double Holliday Junction Dissolution Mediated by the BLM-Topo III?-RMI1-RMI2 Protein Complex *
    Article Snippet: The supernatant, wash, and SDS eluate, 10 μl of each, were analyzed by 7.5% SDS-PAGE and Coomassie Blue staining. .. For the affinity pull-down of MBP-tagged proteins, 5 μg of hRPA was incubated with MBP-tagged RMI1425–625 -RMI2, RMI1-RMI2, RMI1Δ301–337 -RMI2, RMI14EA -RMI2, or MBP (5 μg) and mixed with amylose resin (New England Biolabs) to capture the MBP-tagged protein and associated RPA, as above.

    Article Title: The Transcription Factor Atf1 Binds and Activates the APC/C Ubiquitin Ligase in Fission Yeast *
    Article Snippet: S. pombe MBP-tagged E1 (Uba1) and His-tagged E2s (Ubc1, Ubc4, and Ubc11) were expressed in Escherichia coli and purified using amylose resin (New England Biolab) or Ni-NTA beads (Qiagen) as described by the supplier. .. Reactions were stopped at the indicated time points with SDS sample buffer, and mixtures were resolved by SDS-PAGE with detection by fluorography.

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin
    Article Snippet: Sepharose was washed and bound proteins were analyzed by 12.5% SDS–PAGE and Coomassie blue staining. .. Bcl-2-binding assay : A 40 μl volume of 6 μM MBP-Bcl-2 fusion protein (MBP-Bcl-2) was subjected to 40 μl amylose resin (New England Biolabs, Beverly, MA) and incubated for 30 min.

    Article Title: Protein domain definition should allow for conditional disorder
    Article Snippet: The MBP fusion proteins were isolated using a bed of amylose resin (New England Biolabs), and eluted in lysis buffer supplemented with 30 m M maltose. .. Protein purity was assessed using SDS-PAGE, coupled with colloidal Coomassie staining for protein detection.

    Article Title: Stepwise Activity of ClpY (HslU) Mutants in the Processive Degradation of Escherichia coli ClpYQ (HslUV) Protease Substrates ▿
    Article Snippet: ClpY and its above derivative mutants were each purified using His tag affinity purification methods, whereas MBP-SulA was purified using amylose resin (NEB) methods and the concentration of each protein was determined (see Materials and Methods). .. As shown in , each of the various ClpY mutants was capable of pulling down with MBP-SulA, seen in the SDS-PAGE gel and in the Western blot analyses.

    Article Title: A general path for large-scale solubilization of cellular proteins: From membrane receptors to multiprotein complexes
    Article Snippet: We have used the following affinity beads with high success: (1) nickel beads for poly-His tagged proteins (Sigma); (2) antibody beads (IgG, GE healthcare) for protein-A-tagged (PA-tag) proteins; (3) amylose resin (New England Biolabs) for proteins fused to maltose binding protein tag and, (4) chitin beads for proteins purified using the IMPACT-twin purification system (New England Biolabs). .. Following incubation, beads are thoroughly washed several times with buffer A plus 2.5× CMC of the corresponding detergent, and eluted (or boiled); individual fractions are subject to SDS–PAGE to quantify protein extraction and quality of purification.

    Article Title: Molecular Determinants of Gem Protein Inhibition of P/Q-type Ca2+ Channels *
    Article Snippet: MBP fusion peptides were expressed in DE3 and purified by incubation with amylose resin (New England Biolabs). .. The elution was detected with Coomassie Blue staining on SDS-PAGE gel.

    Plasmid Preparation:

    Article Title: Quantitation of SPLUNC1 in saliva with an xMAP particle-based antibody capture and detection immunoassay
    Article Snippet: Briefly, the cDNA for human SPLUNC1 (National Center for Biotechnology Information, NCBI accession number NM 016583) was cloned into the plasmid vector pMAL-c2x (New England Biolabs, Ipswich, MA, USA) for the expression of a fusion protein containing an N-terminal maltose-binding protein tag and a C-terminal 6xHis tag. .. Full-length fusion protein was expressed in the Escherichia coli strain BL21 Star (DE3, Gibco, Invitrogen Corp., Carlsbad, CA, USA). rhSPLUNC1 was purified first, by passing the fusion protein preparation through a column containing amylose resin (New England Biolabs) and then, by passing the fraction containing rhSPLUNC1 through a nickel resin column (Ni Sepharose 6 Fast Flow, GE Healthcare Biosciences Corp., Piscataway, NJ, USA).

    Article Title: An Antibody Directed against the Fusion Peptide of Jun?n Virus Envelope Glycoprotein GPC Inhibits pH-Induced Membrane Fusion ▿
    Article Snippet: The G2 ectodomain region of the molecularly cloned JUNV GPC (amino acids 252 to 424) was appended in frame to the C terminus of maltose-binding protein (MBP) in the pMAL-c2E vector (New England Biolabs) using standard PCR and molecular cloning techniques. .. Aliquots of the cleared lysate were incubated with a 50% slurry of amylose resin (New England BioLabs) for 2 h at 4°C to purify the MBP fusion proteins.

    SYBR Green Assay:

    Article Title: Structural Basis for Influenza Virus NS1 Protein Block of mRNA Nuclear Export
    Article Snippet: .. 50xAdvantage Polymerase mix (Clontech (EMD), 639202); dNTP’s (Clontech (EMD), 639125); BamHI-HF (New England Biolabs (NEB), R3136T); NotI (New England Biolabs (NEB), R0189S); SmaI (New England Biolabs (NEB), R0141); XhoI (New England Biolabs (NEB), R0146S); T4 DNA Ligase (New England Biolabs (NEB), M0202); QIAquick Gel Extraction Kit (Qiagen, 28704); NEB® 5-alpha Competent E. coli (New England Biolabs (NEB), ); Rosetta (DE3) Competent Cells (EMD Millipore, 70954); SOC Outgrowth Medium (New England Biolabs (NEB), B9020); QIAprep Spin Miniprep Kit (250) (Qiagen, 27106); Lipofectamine® RNAiMAX Transfection Reagent (Thermo Fisher Scientific, 13778150); RNeasy Plus Mini Kit (Qiagen, 74134); Random Hexamers (50 μM) (Thermo Fisher Scientific, N8080127); Protector RNase Inhibitor (Roche, 03335402001); SuperScript II Reverse Transcriptase (Thermo Fisher Scientific, 18064014); LIGHTCYCLER 480 SYBR GREEN I MASTER (Roche, 04707516001); LightCycler® 480 Multiwell Plate 96, White (Roche, 04729692001); NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, 78833); Complete EDTA-free protease inhibitor tablets (Sigma-Aldrich, 11873580001); TransIT-X2® Dynamic Delivery System (Mirus Bio, MIR6000); SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific, 34096); L-Glutathione reduced (Sigma-Aldrich, G4251–25G); Ampicillin (Sigma-Aldrich, A-9518); Kanamycin Monosulfate (Gold Biotechnology, K-120–10); IPTG (Gold Biotechnology, I2481C50); Imidazole (Sigma-Aldrich, 56750); PMSF (RPI, ); Aprotinin (Santa Cruz Biotechnology, sc-3595); Leupeptin (Santa Cruz Biotechnology, sc295358); Pepstatin A (Thermo Fisher Scientific, ); Glutathione Sepharose 4B (GE Healthcare, 17–0756-01); Amylose Resin (New England Biolabs (NEB), E8021S); Ni-NTA Agarose (Qiagen, 30210); Mono Q 5/50 GL (GE Healthcare, 17-5166-01); HiTrap SP HP (GE Healthcare, 17-1151-01); Superdex 200 HR 10/30 (GE Healthcare, 17-1088-01); EasyTag EXPRESS35 S Protein Labeling Mix, [35S]-, 7mCi (PerkinElmer, NEG772007MC); T7 RiboMAX™ Express Large Scale RNA Production System (Promega, P1320); Protein G Sepharose® 4 Fast Flow (GE Healthcare, 17-0618-01); Hoechst 33258 (Molecular Probes/Life Technologies); M mRNA probes (Biosearch Technologies) ; ProLong Gold antifade reagent (Life Technologies, ). ..

    Binding Assay:

    Article Title: Resistance protein Pit interacts with the GEF OsSPK1 to activate OsRac1 and trigger rice immunity
    Article Snippet: Paragraph title: In Vitro Binding Assay. ... The mixture was applied to 30 µL Amylose resin (New England Biolabs) or Glutathione Sepharose 4B (GE Healthcare) and incubated at 4 °C for 2–3 h. The beads were then washed five times for 5 min each time with pull-down buffer, and the bound proteins were eluted with 100 µl 2× SDS loading buffer and subjected to immunoblot assay with anti-SUMO (A01693; GenScript), anti-GST (sc-138; Santa Cruz Biotechnology), and anti-MBP (E8032S; New England Biolabs) antibodies.

    Article Title: Dual DNA-binding domains shape the interaction of Brh2 with DNA
    Article Snippet: Brh2NT terminating at residue M551 was purified as a fusion protein with an N-terminal maltose binding protein (MBP)-tag and C-terminal hexahistidine (His)-tag after expression in E. coli . .. Purification involved sequential affinity chromatography on Ni2+ -NTA (nitrilotriacetate) agarose (Qiagen), amylose resin (New England Biolabs), salt gradient elution from monoQ ion exchange resin, and salt gradient elution from HiTrap heparin-agarose using an AKTA FPLC chromatography system (GE Healthcare).

    Article Title: Structure and Mechanism of the Lipooligosaccharide Sialyltransferase from Neisseria meningitidis *
    Article Snippet: .. The lysate was centrifuged at 27,000 × g for 30 min, and the resulting supernatant was loaded onto a column (1 column volume of 20 ml) of amylose resin (New England Biolabs) that was equilibrated with the binding buffer (2 m m EDTA (pH 7.5), 20 m m Tris-HCl (pH 7.5), 200 m m NaCl, 10% (v/v) glycerol, and 5 m m β-mercaptoethanol). .. The bound MBP-Δ29NST fusion protein was fractionated with the elution buffer (binding buffer plus 10 m m maltose).

    Article Title: Protein domain definition should allow for conditional disorder
    Article Snippet: The MBP fusion proteins were isolated using a bed of amylose resin (New England Biolabs), and eluted in lysis buffer supplemented with 30 m M maltose. .. Proteins were further purified using anion exchange chromatography, binding them to Q Sepharose HP resin (Amersham Biosciences) buffered with 12.5 m M Tris/HCl pH 8.5, 50 m M NaCl, and then displacing them with a linear salt gradient.

    Article Title: A general path for large-scale solubilization of cellular proteins: From membrane receptors to multiprotein complexes
    Article Snippet: .. We have used the following affinity beads with high success: (1) nickel beads for poly-His tagged proteins (Sigma); (2) antibody beads (IgG, GE healthcare) for protein-A-tagged (PA-tag) proteins; (3) amylose resin (New England Biolabs) for proteins fused to maltose binding protein tag and, (4) chitin beads for proteins purified using the IMPACT-twin purification system (New England Biolabs). .. Following incubation, beads are thoroughly washed several times with buffer A plus 2.5× CMC of the corresponding detergent, and eluted (or boiled); individual fractions are subject to SDS–PAGE to quantify protein extraction and quality of purification.

    In Vitro:

    Article Title: Resistance protein Pit interacts with the GEF OsSPK1 to activate OsRac1 and trigger rice immunity
    Article Snippet: Paragraph title: In Vitro Binding Assay. ... The mixture was applied to 30 µL Amylose resin (New England Biolabs) or Glutathione Sepharose 4B (GE Healthcare) and incubated at 4 °C for 2–3 h. The beads were then washed five times for 5 min each time with pull-down buffer, and the bound proteins were eluted with 100 µl 2× SDS loading buffer and subjected to immunoblot assay with anti-SUMO (A01693; GenScript), anti-GST (sc-138; Santa Cruz Biotechnology), and anti-MBP (E8032S; New England Biolabs) antibodies.

    Article Title: Stepwise Activity of ClpY (HslU) Mutants in the Processive Degradation of Escherichia coli ClpYQ (HslUV) Protease Substrates ▿
    Article Snippet: Paragraph title: In vitro protein pulldown analyses of ClpY and its derivative mutants with MBP-SulA. ... ClpY and its above derivative mutants were each purified using His tag affinity purification methods, whereas MBP-SulA was purified using amylose resin (NEB) methods and the concentration of each protein was determined (see Materials and Methods).

    Gel Permeation Chromatography:

    Article Title: An Antibody Directed against the Fusion Peptide of Jun?n Virus Envelope Glycoprotein GPC Inhibits pH-Induced Membrane Fusion ▿
    Article Snippet: The G2 ectodomain region of the molecularly cloned JUNV GPC (amino acids 252 to 424) was appended in frame to the C terminus of maltose-binding protein (MBP) in the pMAL-c2E vector (New England Biolabs) using standard PCR and molecular cloning techniques. .. Aliquots of the cleared lysate were incubated with a 50% slurry of amylose resin (New England BioLabs) for 2 h at 4°C to purify the MBP fusion proteins.

    Produced:

    Article Title: The Transcription Factor Atf1 Binds and Activates the APC/C Ubiquitin Ligase in Fission Yeast *
    Article Snippet: Substrates (Cdc13 and Cut2) and activator Ste9/Srw1 were produced by coupled transcription/translation in reticulocyte lysate (T n T; Promega) from the plasmids. .. S. pombe MBP-tagged E1 (Uba1) and His-tagged E2s (Ubc1, Ubc4, and Ubc11) were expressed in Escherichia coli and purified using amylose resin (New England Biolab) or Ni-NTA beads (Qiagen) as described by the supplier.

    Concentration Assay:

    Article Title: Stepwise Activity of ClpY (HslU) Mutants in the Processive Degradation of Escherichia coli ClpYQ (HslUV) Protease Substrates ▿
    Article Snippet: .. ClpY and its above derivative mutants were each purified using His tag affinity purification methods, whereas MBP-SulA was purified using amylose resin (NEB) methods and the concentration of each protein was determined (see Materials and Methods). .. Each ClpY mutant protein and the wild-type ClpY were equivalently tested for their own pulling down with MBP-SulA in the presence of ATP.

    Lysis:

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin
    Article Snippet: Bcl-2-binding assay : A 40 μl volume of 6 μM MBP-Bcl-2 fusion protein (MBP-Bcl-2) was subjected to 40 μl amylose resin (New England Biolabs, Beverly, MA) and incubated for 30 min. .. To prepare SH-SY5Y crude cell extract, cells were harvested, centrifuged for 10 min at 2000 g and cell pellets were resuspended in the same volume of hypotonic lysis buffer, followed by sonication.

    Article Title: Structure and Mechanism of the Lipooligosaccharide Sialyltransferase from Neisseria meningitidis *
    Article Snippet: The cell pellet was resuspended in lysis buffer (2 m m EDTA (pH 7.5), 20 m m Tris-HCl (pH 7.5), 200 m m NaCl, 10% (v/v) glycerol) supplemented with a Roche Applied Science EDTA-free protease inhibitor mixture tablet and lysed at 20,000 p.s.i. using the Avestin Emulsiflex homogenizer. .. The lysate was centrifuged at 27,000 × g for 30 min, and the resulting supernatant was loaded onto a column (1 column volume of 20 ml) of amylose resin (New England Biolabs) that was equilibrated with the binding buffer (2 m m EDTA (pH 7.5), 20 m m Tris-HCl (pH 7.5), 200 m m NaCl, 10% (v/v) glycerol, and 5 m m β-mercaptoethanol).

    Article Title: Protein domain definition should allow for conditional disorder
    Article Snippet: .. The MBP fusion proteins were isolated using a bed of amylose resin (New England Biolabs), and eluted in lysis buffer supplemented with 30 m M maltose. .. Proteins were further purified using anion exchange chromatography, binding them to Q Sepharose HP resin (Amersham Biosciences) buffered with 12.5 m M Tris/HCl pH 8.5, 50 m M NaCl, and then displacing them with a linear salt gradient.

    Gel Extraction:

    Article Title: Structural Basis for Influenza Virus NS1 Protein Block of mRNA Nuclear Export
    Article Snippet: .. 50xAdvantage Polymerase mix (Clontech (EMD), 639202); dNTP’s (Clontech (EMD), 639125); BamHI-HF (New England Biolabs (NEB), R3136T); NotI (New England Biolabs (NEB), R0189S); SmaI (New England Biolabs (NEB), R0141); XhoI (New England Biolabs (NEB), R0146S); T4 DNA Ligase (New England Biolabs (NEB), M0202); QIAquick Gel Extraction Kit (Qiagen, 28704); NEB® 5-alpha Competent E. coli (New England Biolabs (NEB), ); Rosetta (DE3) Competent Cells (EMD Millipore, 70954); SOC Outgrowth Medium (New England Biolabs (NEB), B9020); QIAprep Spin Miniprep Kit (250) (Qiagen, 27106); Lipofectamine® RNAiMAX Transfection Reagent (Thermo Fisher Scientific, 13778150); RNeasy Plus Mini Kit (Qiagen, 74134); Random Hexamers (50 μM) (Thermo Fisher Scientific, N8080127); Protector RNase Inhibitor (Roche, 03335402001); SuperScript II Reverse Transcriptase (Thermo Fisher Scientific, 18064014); LIGHTCYCLER 480 SYBR GREEN I MASTER (Roche, 04707516001); LightCycler® 480 Multiwell Plate 96, White (Roche, 04729692001); NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, 78833); Complete EDTA-free protease inhibitor tablets (Sigma-Aldrich, 11873580001); TransIT-X2® Dynamic Delivery System (Mirus Bio, MIR6000); SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific, 34096); L-Glutathione reduced (Sigma-Aldrich, G4251–25G); Ampicillin (Sigma-Aldrich, A-9518); Kanamycin Monosulfate (Gold Biotechnology, K-120–10); IPTG (Gold Biotechnology, I2481C50); Imidazole (Sigma-Aldrich, 56750); PMSF (RPI, ); Aprotinin (Santa Cruz Biotechnology, sc-3595); Leupeptin (Santa Cruz Biotechnology, sc295358); Pepstatin A (Thermo Fisher Scientific, ); Glutathione Sepharose 4B (GE Healthcare, 17–0756-01); Amylose Resin (New England Biolabs (NEB), E8021S); Ni-NTA Agarose (Qiagen, 30210); Mono Q 5/50 GL (GE Healthcare, 17-5166-01); HiTrap SP HP (GE Healthcare, 17-1151-01); Superdex 200 HR 10/30 (GE Healthcare, 17-1088-01); EasyTag EXPRESS35 S Protein Labeling Mix, [35S]-, 7mCi (PerkinElmer, NEG772007MC); T7 RiboMAX™ Express Large Scale RNA Production System (Promega, P1320); Protein G Sepharose® 4 Fast Flow (GE Healthcare, 17-0618-01); Hoechst 33258 (Molecular Probes/Life Technologies); M mRNA probes (Biosearch Technologies) ; ProLong Gold antifade reagent (Life Technologies, ). ..

    Chick Chorioallantoic Membrane Assay:

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin
    Article Snippet: Subsequently, 30 μg recombinant FKBP38 was incubated with CaM-Sepharose. .. Bcl-2-binding assay : A 40 μl volume of 6 μM MBP-Bcl-2 fusion protein (MBP-Bcl-2) was subjected to 40 μl amylose resin (New England Biolabs, Beverly, MA) and incubated for 30 min.

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  • 90
    New England Biolabs amylose resin
    Amylose Resin, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 736 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/amylose resin/product/New England Biolabs
    Average 90 stars, based on 736 article reviews
    Price from $9.99 to $1999.99
    amylose resin - by Bioz Stars, 2020-01
    90/100 stars
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