amylose resin  (New England Biolabs)


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  • 99
    Name:
    Amylose Resin
    Description:
    Amylose Resin 100 ml
    Catalog Number:
    E8021L
    Price:
    1103
    Size:
    100 ml
    Category:
    Protein Purification Kit Components
    Score:
    85
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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 99 stars, based on 198 article reviews
    Price from $9.99 to $1999.99
    amylose resin - by Bioz Stars, 2019-12
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Autophosphorylation of Ser-6 via an intermolecular mechanism is important for the rapid reduction of NtCDPK1 kinase activity for substrate RSG
    Article Snippet: PCR products were cloned into pGEX-4T-1 (GE Healthcare). .. Glutathione S -transferase (GST)-NtCDPK1s, maltose-binding protein (MBP)-RSG, and 10×His-tagged (His)-NtCDPK1 were expressed in Escherichia coli harbouring pGEX-4T-1-NtCDPK1, pMALc2-RSG, or pET16b-NtCDPK1, respectively, and purified by glutathione-Sepharose 4B (GE Healthcare), Amylose Resin (New England Biolabs), or COSMOGEL His-Accept (Nacalai Tesque), respectively.

    Article Title: Role of Tomato Lipoxygenase D in Wound-Induced Jasmonate Biosynthesis and Plant Immunity to Insect Herbivores
    Article Snippet: For plasmid construction of maltose binding protein (MBP) fusions with SlMYC2, the cDNA was amplified and cloned into the pMAL-c2 vector (New England Biolabs, Beverly, MA) via Bam HI and Pst I restriction sites. .. The MBP-SlMYC2 recombinant protein was expressed in the BL21 Escheichia coli (E. coli ) strain and purified by binding onto an amylose resin (New England Biolabs) column, according to the instructions provided by the manufacturer.

    Article Title: DER containing two consecutive GTP-binding domains plays an essential role in chloroplast ribosomal RNA processing and ribosome biogenesis in higher plants
    Article Snippet: To purify recombinant proteins of NbDER and its mutants for GTPase assays, the corresponding NbDER cDNA fragments were PCR-amplified and cloned into the pMALTM c2 or pMAL-c2 vector (New England Biolabs). .. The MBP fusion proteins were purified using amylose resin following the manufacturer’s instructions (New England Biolabs).

    Article Title: The two TRAPP complexes of metazoans have distinct roles and act on different Rab GTPases
    Article Snippet: To purify TRAPPC9 antisera, the N terminus of TRAPPC9 corresponding with amino acids 23–514 of GenBank accession number was fused at its N terminus to His-MBP by cloning the corresponding region of the coding sequence of cDNA clone RE66325 into the expression vector pETM-41. .. The TRAPPC9 (aa23–514)-6×His-MBP fusion protein was purified from the cleared lysate with amylose resin (E8021S; New England Biolabs, Inc.), and coupled with CNBr-activated Sepharose 4B beads.

    Article Title: Identification of multiple binding sites for the THAP domain of the Galileo transposase in the long terminal inverted-repeats
    Article Snippet: Since no ancestral sequence was reconstructed for the D. buzzatii domain, the 450 bp THAP sequence (150 amino acid) was cloned directly in the pOPINM expression vector. .. The supernatant was loaded onto an amylose resin column (New England Biolabs).

    Centrifugation:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation. .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Article Title: WRB and CAML Are Necessary and Sufficient to Mediate Tail-Anchored Protein Targeting to the ER Membrane
    Article Snippet: After lysis with Avestin Emulsiflex-C5 aggregates were removed by centrifugation at 100,000 g , 30 minutes, 4°C. .. The supernatant was loaded onto an amylose resin (NEB) column.

    Article Title: A Cell Cycle and Nutritional Checkpoint Controlling Bacterial Surface Adhesion
    Article Snippet: Cells were disrupted by three passages in a French pressure cell, and cell debris was removed by centrifugation for 20 min (25,000 g, 4°C). .. The supernatant was then mixed with 500 µl of Amylose resin (New England Biolabs) pre-equilibrated with Tris-NaCl buffer, which allowed for binding of MBP domains.

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: After the collection of cells by centrifugation (5,000 rpm for 5 min), the cells were resuspended and sonicated in low-salt column buffer (30 mM HEPES-KOH pH 7.4, 25 mM NaCl, 1 mM EDTA, 10 mM β-mercaptoethanol). .. To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C.

    Article Title: Identification of multiple binding sites for the THAP domain of the Galileo transposase in the long terminal inverted-repeats
    Article Snippet: The cells were harvested by centrifugation and resuspended in HSG buffer, which contained 50 mM HEPES pH 7.5, 200 mM NaCl, 2 mM dithiothreitol, 5 mM EDTA and 10% glycerol. .. The supernatant was loaded onto an amylose resin column (New England Biolabs).

    Article Title: Structure Based Identification and Characterization of Flavonoids That Disrupt Human Papillomavirus-16 E6 Function
    Article Snippet: Cells were lysed in 50 mM Tris HCl pH 6.8, 300 mM NaCl, 0.1 % (v/v) NP-40, 2 mM DTT and 1x protease inhibitor cocktail (EDTA-free, Roche) and sonicated as described before. .. Cell lysate was cleared by centrifugation at 15000xg, 4°C for 30 min and equilibrated with amylose resin beads (New England Biolabs, Inc.) by inversion for 3 hrs at 4°C. .. Bound beads were washed one time in lysis buffer and four times with wash buffer (50 mM Tris-HCl pH 6.8, 400 mM NaCl, 2 mM DTT, 1x protease inhibitor cocktail).

    Amplification:

    Article Title: Role of Tomato Lipoxygenase D in Wound-Induced Jasmonate Biosynthesis and Plant Immunity to Insect Herbivores
    Article Snippet: For plasmid construction of maltose binding protein (MBP) fusions with SlMYC2, the cDNA was amplified and cloned into the pMAL-c2 vector (New England Biolabs, Beverly, MA) via Bam HI and Pst I restriction sites. .. The MBP-SlMYC2 recombinant protein was expressed in the BL21 Escheichia coli (E. coli ) strain and purified by binding onto an amylose resin (New England Biolabs) column, according to the instructions provided by the manufacturer.

    Article Title: DER containing two consecutive GTP-binding domains plays an essential role in chloroplast ribosomal RNA processing and ribosome biogenesis in higher plants
    Article Snippet: The MBP fusion proteins were purified using amylose resin following the manufacturer’s instructions (New England Biolabs). .. Purified proteins of MBP:NbDER, MBP:PM1, MBP:PM2, and MBP:PM1/2 were concentrated using Amicon Ultra Centrifugal Filters (Millipore).

    Article Title: Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases
    Article Snippet: Other materials were provided by the following suppliers: SMART RACE™ cDNA Amplification kit and Advantage 2 PCR System were from CLONTECH, Mountain View, California, USA. .. ; restriction enzymes were from Promega, Chilworth, Southampton, U.K.; chelating Sepharose fast flow resin and gel filtration columns were from GE Healthcare, Little Chalfont, Bucks, U.K.; pET14b was from Novagen, Madison, WI, USA; pMalC2 and amylose resin were from New England Biolabs, Beverly, Maine, USA: all RNA preservation systems were from AMBION, Austin, Texas, USA; RNA extraction kits were from QIAGEN, Crawley, West Sussex, UK; and primers were from Invitrogen, Paisley, UK.

    Filtration:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: For the time course and titration experiments, the Ni2+ -NTA eluate was polished by gel filtration chromatography on a HiLoad 26/60 Superdex 200 prep-grade column equilibrated in standard assay buffer (10 mm HEPES-Na+ , 1 mm EDTA-Na+ , 50 mm NaCl, pH 7.4). .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Article Title: Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases
    Article Snippet: Other materials were provided by the following suppliers: SMART RACE™ cDNA Amplification kit and Advantage 2 PCR System were from CLONTECH, Mountain View, California, USA. .. ; restriction enzymes were from Promega, Chilworth, Southampton, U.K.; chelating Sepharose fast flow resin and gel filtration columns were from GE Healthcare, Little Chalfont, Bucks, U.K.; pET14b was from Novagen, Madison, WI, USA; pMalC2 and amylose resin were from New England Biolabs, Beverly, Maine, USA: all RNA preservation systems were from AMBION, Austin, Texas, USA; RNA extraction kits were from QIAGEN, Crawley, West Sussex, UK; and primers were from Invitrogen, Paisley, UK. .. Coryphaenoides armatus (rat tail) specimens were caught by otter trawl in the Atlantic from the Porcupine Seabight from a depth of 3986 m – 4016 m, September 2000, on RRS Discovery Cruise 250.

    Binding Assay:

    Article Title: Role of Tomato Lipoxygenase D in Wound-Induced Jasmonate Biosynthesis and Plant Immunity to Insect Herbivores
    Article Snippet: For plasmid construction of maltose binding protein (MBP) fusions with SlMYC2, the cDNA was amplified and cloned into the pMAL-c2 vector (New England Biolabs, Beverly, MA) via Bam HI and Pst I restriction sites. .. The MBP-SlMYC2 recombinant protein was expressed in the BL21 Escheichia coli (E. coli ) strain and purified by binding onto an amylose resin (New England Biolabs) column, according to the instructions provided by the manufacturer. .. The 50-bp TomLoxD promoter probes containing G-box-like motif at the -369 site were synthesized and labeled with biotin at the 3′ end (Invitrogen), which containing the same sequences as that of the competitor probes without biotin-labled, while the mutated labeled probes were deleted the G-box-like motif.

    Article Title: A Cell Cycle and Nutritional Checkpoint Controlling Bacterial Surface Adhesion
    Article Snippet: Cells were disrupted by three passages in a French pressure cell, and cell debris was removed by centrifugation for 20 min (25,000 g, 4°C). .. The supernatant was then mixed with 500 µl of Amylose resin (New England Biolabs) pre-equilibrated with Tris-NaCl buffer, which allowed for binding of MBP domains. .. The resin was thoroughly washed with the Tris-NaCl buffer and bound proteins were eluted with 500 µl of Tris-NaCl buffer supplemented with 20 mM maltose.

    Article Title: The Self-Interaction of a Nodavirus Replicase Is Enhanced by Mitochondrial Membrane Lipids
    Article Snippet: Cell pellets were resuspended in binding buffer (20 mM Tris-HCl [pH 7.4], 200 mM NaCl, 1 mM EDTA, 10 mM 2-Mercaptoethanol) supplemented with 1.5% Triton-X 100 and protease inhibitors cocktail (Sigma, St. Louis, Mo, USA). .. The proteins in the supernatant were purified using amylose resin (New England BioLabs) according to the manufacturer’s protocol and concentrated using Amicon Ultra-15 filters (Millipore, Schwalbach, Germany), and the buffer was exchanged to the hypotonic buffer (1 mM HEPES [pH 7.4], 0.1 mM EDTA, 15 mM NaCl, 1 mM DTT).

    Article Title: Novel interaction of Rab13 and Rab8 with endospanins
    Article Snippet: Cells were suspended in binding buffer (50 mM Tris, 300 mM NaCl, 5 mM MgCl2 , 1 mM EDTA, 10 mM β-mercaptoethanol, 0.1% Triton X-100 and protease inhibitors (Roche), pH 7.2) and lysed by sonication. .. Equimolar amounts of MBP-bait proteins (∼10 μg) were incubated with GST-target proteins (∼2 μg) for 2 h at 4 °C together with amylose resin beads (New England BioLabs).

    Construct:

    Article Title: Structure of CbpA J-Domain Bound to the Regulatory Protein CbpM Explains Its Specificity and Suggests Evolutionary Link between CbpM and Transcriptional Regulators
    Article Snippet: The tags were cleaved using either thrombin or tobacco-etch virus protease (TEV), depending on the construct. .. For CbpM and CbpA, the protein samples were also passed through amylose resin (New England Biolabs Inc).

    SDS-Gel:

    Article Title: C-Terminal Domain of ICA69 Interacts with PICK1 and Acts on Trafficking of PICK1-PKC? Complex and Cerebellar Plasticity
    Article Snippet: Amylose resin (New England Biolabs, Hitchin, UK) was added to cleared lysates. .. Fusion protein concentration was determined using a BCA kit assay (Thermo).

    Incubation:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: The protein concentration was determined using a Nanodrop 1000 (Thermo Scientific) and the concentrated protein was stored at 4 °C for up to 2 weeks without a noticeable decrease in activity. .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation. .. The resin was poured into a 1.5 × 12-cm chromatography column (Bio-Rad), washed with ∼100 column volumes of MBP buffer (20 mm Tris-HCl, pH 7.4, 0.2 m NaCl), and bound protein was eluted with 10 ml of MBP buffer containing 50 mm maltose.

    Article Title: C-Terminal Domain of ICA69 Interacts with PICK1 and Acts on Trafficking of PICK1-PKC? Complex and Cerebellar Plasticity
    Article Snippet: Pellets were lysed in STE buffer (10 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, pH 8.0) containing 1 mg/ml lysozyme and incubated with rotation at 4°C for 1 hr. .. Amylose resin (New England Biolabs, Hitchin, UK) was added to cleared lysates.

    Article Title: Mitotic spindle scaling during Xenopus development by kif2a and importin ?
    Article Snippet: The NLS mutant had three charge switch mutations, K427E, K428E, and K430E, introduced into the NLS sequence identified by Quikchange mutagenesis (Stratagene/Agilent Technologies, Santa Clara CA, USA). .. 1 µM recombinant kif2a or kif2a ΔNLS as an MBP fusion protein or MBP alone was added to extracts and incubated for 15 min. 75 µl of Amylose resin (New England Biolabs) was then incubated for 15 min to retrieved MBP or fusion proteins. .. Resin was washed with a total of 25 bed volumes of phosphate-buffered saline with 0.01% Tween and eluted in SDS-PAGE loading buffer.

    Article Title: WRB and CAML Are Necessary and Sufficient to Mediate Tail-Anchored Protein Targeting to the ER Membrane
    Article Snippet: The supernatant was loaded onto an amylose resin (NEB) column. .. The supernatant was loaded onto an amylose resin (NEB) column.

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: After the collection of cells by centrifugation (5,000 rpm for 5 min), the cells were resuspended and sonicated in low-salt column buffer (30 mM HEPES-KOH pH 7.4, 25 mM NaCl, 1 mM EDTA, 10 mM β-mercaptoethanol). .. To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C. .. After careful washing of the columns, the proteins were eluted with MBP-elution buffer [column buffer containing 0.18% (W/V) maltose].

    Article Title: Cep63 and Cep152 Cooperate to Ensure Centriole Duplication
    Article Snippet: After washing in column buffer, amylose resin was incubated with 1 mg cell lysate from cells 293 expressing GFP-Cep152 full length, N-, or C-terminal domains, in lysis buffer for 2 hours at 4°C. .. MBP and MBP-Cep63 were expressed in BL21 CodonPlus (DE3) RIL E. coli (Stratagene) at 16°C for 3 hours after induction with 0.3 mM IPTG (Isopropyl β-D-1-thiogalactopyranoside), and purified using amylose resin according the manufacturer’s protocol (NEB).

    Article Title: 18S rRNA processing requires base pairings of snR30 H/ACA snoRNA to eukaryote-specific 18S sequences
    Article Snippet: The spheroplast suspension was transferred onto ice, supplemented with 100 μg/ml AMT psoralen (4′-aminomethyl-4,5′,8-trimethylpsoralen) (HRI Associates Inc.), incubated in the dark for 15 min and irradiated with UV light (365 nm, ∼25 mW/cm2 ) for 15 min. Total RNA was extracted from the irradiated spheroplasts. .. For affinity selection of M2-R30 RNA, about 20 μg of purified recombinant MS2-MBP protein was bound to 20 μl of amylose resin (New England BioLabs) in NET2 buffer (200 mM NaCl, 0.05% Nonidet P-40 and 20 mM Tris–HCl pH 7.4) as described ( ).

    Article Title: Novel interaction of Rab13 and Rab8 with endospanins
    Article Snippet: Cells were suspended in binding buffer (50 mM Tris, 300 mM NaCl, 5 mM MgCl2 , 1 mM EDTA, 10 mM β-mercaptoethanol, 0.1% Triton X-100 and protease inhibitors (Roche), pH 7.2) and lysed by sonication. .. Equimolar amounts of MBP-bait proteins (∼10 μg) were incubated with GST-target proteins (∼2 μg) for 2 h at 4 °C together with amylose resin beads (New England BioLabs). .. The beads were washed, and bound proteins were eluted in 10 mM maltose and processed for western blotting.

    Activity Assay:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: The protein concentration was determined using a Nanodrop 1000 (Thermo Scientific) and the concentrated protein was stored at 4 °C for up to 2 weeks without a noticeable decrease in activity. .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Expressing:

    Article Title: WRB and CAML Are Necessary and Sufficient to Mediate Tail-Anchored Protein Targeting to the ER Membrane
    Article Snippet: Paragraph title: Protein Expression and Purification ... The supernatant was loaded onto an amylose resin (NEB) column.

    Article Title: A Cell Cycle and Nutritional Checkpoint Controlling Bacterial Surface Adhesion
    Article Snippet: When the culture reached OD660 ∼0.8, 0.5 mM of IPTG was added to induce expression. .. The supernatant was then mixed with 500 µl of Amylose resin (New England Biolabs) pre-equilibrated with Tris-NaCl buffer, which allowed for binding of MBP domains.

    Article Title: The Self-Interaction of a Nodavirus Replicase Is Enhanced by Mitochondrial Membrane Lipids
    Article Snippet: The expression and purification of recombinant WhNV protein A and its derivatives were carried out as previously described – , . .. The proteins in the supernatant were purified using amylose resin (New England BioLabs) according to the manufacturer’s protocol and concentrated using Amicon Ultra-15 filters (Millipore, Schwalbach, Germany), and the buffer was exchanged to the hypotonic buffer (1 mM HEPES [pH 7.4], 0.1 mM EDTA, 15 mM NaCl, 1 mM DTT).

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: Protein expression of the selected helicase proteins was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) for 8 h at 23°C and in the case of viral proteins p33 and p92 at 16°C. .. To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C.

    Article Title: Structure of CbpA J-Domain Bound to the Regulatory Protein CbpM Explains Its Specificity and Suggests Evolutionary Link between CbpM and Transcriptional Regulators
    Article Snippet: Paragraph title: Protein Expression and Purification ... For CbpM and CbpA, the protein samples were also passed through amylose resin (New England Biolabs Inc).

    Article Title: Cep63 and Cep152 Cooperate to Ensure Centriole Duplication
    Article Snippet: After washing in column buffer, amylose resin was incubated with 1 mg cell lysate from cells 293 expressing GFP-Cep152 full length, N-, or C-terminal domains, in lysis buffer for 2 hours at 4°C. .. MBP and MBP-Cep63 were expressed in BL21 CodonPlus (DE3) RIL E. coli (Stratagene) at 16°C for 3 hours after induction with 0.3 mM IPTG (Isopropyl β-D-1-thiogalactopyranoside), and purified using amylose resin according the manufacturer’s protocol (NEB).

    Article Title: 18S rRNA processing requires base pairings of snR30 H/ACA snoRNA to eukaryote-specific 18S sequences
    Article Snippet: Yeast GAL∷snR30 cells carrying the pMS2-R30 expression plasmid were grown in YPD liquid medium containing 2% sucrose at 30°C until OD600 0.8. .. For affinity selection of M2-R30 RNA, about 20 μg of purified recombinant MS2-MBP protein was bound to 20 μl of amylose resin (New England BioLabs) in NET2 buffer (200 mM NaCl, 0.05% Nonidet P-40 and 20 mM Tris–HCl pH 7.4) as described ( ).

    Article Title: The two TRAPP complexes of metazoans have distinct roles and act on different Rab GTPases
    Article Snippet: The expression of this protein was induced overnight at 16°C in E. coli BL21 (DE3) with 0.2 mM IPTG and 0.2% glucose. .. The TRAPPC9 (aa23–514)-6×His-MBP fusion protein was purified from the cleared lysate with amylose resin (E8021S; New England Biolabs, Inc.), and coupled with CNBr-activated Sepharose 4B beads.

    Article Title: Identification of multiple binding sites for the THAP domain of the Galileo transposase in the long terminal inverted-repeats
    Article Snippet: Paragraph title: THAP protein expression ... The supernatant was loaded onto an amylose resin column (New England Biolabs).

    Article Title: Structure Based Identification and Characterization of Flavonoids That Disrupt Human Papillomavirus-16 E6 Function
    Article Snippet: Protein expression was performed by inoculation of a 1 mL starter culture in 100 mL of Turbo-Broth containing 0.2 % lactose, 0.05% glucose and 0.6% glycerol at 20°C for 24 hrs. .. Cell lysate was cleared by centrifugation at 15000xg, 4°C for 30 min and equilibrated with amylose resin beads (New England Biolabs, Inc.) by inversion for 3 hrs at 4°C.

    Article Title: Novel interaction of Rab13 and Rab8 with endospanins
    Article Snippet: 2.5 MBP- or GST-fusion proteins were produced by bacterial expression and cellular pellets were collected. .. Equimolar amounts of MBP-bait proteins (∼10 μg) were incubated with GST-target proteins (∼2 μg) for 2 h at 4 °C together with amylose resin beads (New England BioLabs).

    BIA-KA:

    Article Title: C-Terminal Domain of ICA69 Interacts with PICK1 and Acts on Trafficking of PICK1-PKC? Complex and Cerebellar Plasticity
    Article Snippet: Amylose resin (New England Biolabs, Hitchin, UK) was added to cleared lysates. .. Purified MBP fusion proteins were then dialyzed against internal solution.

    Article Title: Structure Based Identification and Characterization of Flavonoids That Disrupt Human Papillomavirus-16 E6 Function
    Article Snippet: Cell lysate was cleared by centrifugation at 15000xg, 4°C for 30 min and equilibrated with amylose resin beads (New England Biolabs, Inc.) by inversion for 3 hrs at 4°C. .. Bound MBP-E6 proteins were eluted using a final concentration of 20 mM maltose in 5 bed volumes of wash buffer.

    Western Blot:

    Article Title: Cep63 and Cep152 Cooperate to Ensure Centriole Duplication
    Article Snippet: Paragraph title: Western Blotting, Immuno-precipitations and Pull-down Assays ... MBP and MBP-Cep63 were expressed in BL21 CodonPlus (DE3) RIL E. coli (Stratagene) at 16°C for 3 hours after induction with 0.3 mM IPTG (Isopropyl β-D-1-thiogalactopyranoside), and purified using amylose resin according the manufacturer’s protocol (NEB).

    Article Title: Novel interaction of Rab13 and Rab8 with endospanins
    Article Snippet: Equimolar amounts of MBP-bait proteins (∼10 μg) were incubated with GST-target proteins (∼2 μg) for 2 h at 4 °C together with amylose resin beads (New England BioLabs). .. MBP-fusion proteins were bound to amylose resin beads and incubated with HeLa or Cos1 cell lysates in binding buffer for 2 h at 4 °C.

    Transformation Assay:

    Article Title: C-Terminal Domain of ICA69 Interacts with PICK1 and Acts on Trafficking of PICK1-PKC? Complex and Cerebellar Plasticity
    Article Snippet: MBP, MBP-ICA69, MBP-ICAC, and MBP-ΔICAC were transformed into E. coli BL21 (DE3), respectively. .. Amylose resin (New England Biolabs, Hitchin, UK) was added to cleared lysates.

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: Briefly, the expression plasmids were transformed into E. coli strain BL21 (DE3) CodonPlus. .. To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C.

    Article Title: Identification of multiple binding sites for the THAP domain of the Galileo transposase in the long terminal inverted-repeats
    Article Snippet: The expression vectors with the THAP domains were sequenced to confirm the ORF and transformed in BL21 (DE3) E. coli for protein expression. .. The supernatant was loaded onto an amylose resin column (New England Biolabs).

    Article Title: Structure Based Identification and Characterization of Flavonoids That Disrupt Human Papillomavirus-16 E6 Function
    Article Snippet: MBP-16E6 DNA, a kind gift from G. Travé [ ], was transformed into BL21DE(3) E. coli . .. Cell lysate was cleared by centrifugation at 15000xg, 4°C for 30 min and equilibrated with amylose resin beads (New England Biolabs, Inc.) by inversion for 3 hrs at 4°C.

    RNA Binding Assay:

    Article Title: DER containing two consecutive GTP-binding domains plays an essential role in chloroplast ribosomal RNA processing and ribosome biogenesis in higher plants
    Article Snippet: The MBP fusion proteins were purified using amylose resin following the manufacturer’s instructions (New England Biolabs). .. Purified proteins of MBP:NbDER, MBP:PM1, MBP:PM2, and MBP:PM1/2 were concentrated using Amicon Ultra Centrifugal Filters (Millipore).

    Flow Cytometry:

    Article Title: Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases
    Article Snippet: Other materials were provided by the following suppliers: SMART RACE™ cDNA Amplification kit and Advantage 2 PCR System were from CLONTECH, Mountain View, California, USA. .. ; restriction enzymes were from Promega, Chilworth, Southampton, U.K.; chelating Sepharose fast flow resin and gel filtration columns were from GE Healthcare, Little Chalfont, Bucks, U.K.; pET14b was from Novagen, Madison, WI, USA; pMalC2 and amylose resin were from New England Biolabs, Beverly, Maine, USA: all RNA preservation systems were from AMBION, Austin, Texas, USA; RNA extraction kits were from QIAGEN, Crawley, West Sussex, UK; and primers were from Invitrogen, Paisley, UK. .. Coryphaenoides armatus (rat tail) specimens were caught by otter trawl in the Atlantic from the Porcupine Seabight from a depth of 3986 m – 4016 m, September 2000, on RRS Discovery Cruise 250.

    Chromatography:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: For the time course and titration experiments, the Ni2+ -NTA eluate was polished by gel filtration chromatography on a HiLoad 26/60 Superdex 200 prep-grade column equilibrated in standard assay buffer (10 mm HEPES-Na+ , 1 mm EDTA-Na+ , 50 mm NaCl, pH 7.4). .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Ligation:

    Article Title: Autophosphorylation of Ser-6 via an intermolecular mechanism is important for the rapid reduction of NtCDPK1 kinase activity for substrate RSG
    Article Snippet: NtCDPK1 S6A/D219N and T21A/D219N were generated using overlap extension PCR and seamless ligation cloning extract (SLiCE) [ ] with a plasmid harbouring full-length NtCDPK1 as a template and appropriate primers ( ). .. Glutathione S -transferase (GST)-NtCDPK1s, maltose-binding protein (MBP)-RSG, and 10×His-tagged (His)-NtCDPK1 were expressed in Escherichia coli harbouring pGEX-4T-1-NtCDPK1, pMALc2-RSG, or pET16b-NtCDPK1, respectively, and purified by glutathione-Sepharose 4B (GE Healthcare), Amylose Resin (New England Biolabs), or COSMOGEL His-Accept (Nacalai Tesque), respectively.

    Protease Inhibitor:

    Article Title: Structure Based Identification and Characterization of Flavonoids That Disrupt Human Papillomavirus-16 E6 Function
    Article Snippet: Cells were lysed in 50 mM Tris HCl pH 6.8, 300 mM NaCl, 0.1 % (v/v) NP-40, 2 mM DTT and 1x protease inhibitor cocktail (EDTA-free, Roche) and sonicated as described before. .. Cell lysate was cleared by centrifugation at 15000xg, 4°C for 30 min and equilibrated with amylose resin beads (New England Biolabs, Inc.) by inversion for 3 hrs at 4°C.

    Generated:

    Article Title: Autophosphorylation of Ser-6 via an intermolecular mechanism is important for the rapid reduction of NtCDPK1 kinase activity for substrate RSG
    Article Snippet: NtCDPK1 S6A/D219N and T21A/D219N were generated using overlap extension PCR and seamless ligation cloning extract (SLiCE) [ ] with a plasmid harbouring full-length NtCDPK1 as a template and appropriate primers ( ). .. Glutathione S -transferase (GST)-NtCDPK1s, maltose-binding protein (MBP)-RSG, and 10×His-tagged (His)-NtCDPK1 were expressed in Escherichia coli harbouring pGEX-4T-1-NtCDPK1, pMALc2-RSG, or pET16b-NtCDPK1, respectively, and purified by glutathione-Sepharose 4B (GE Healthcare), Amylose Resin (New England Biolabs), or COSMOGEL His-Accept (Nacalai Tesque), respectively.

    Protein Concentration:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: The protein concentration was determined using a Nanodrop 1000 (Thermo Scientific) and the concentrated protein was stored at 4 °C for up to 2 weeks without a noticeable decrease in activity. .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Article Title: C-Terminal Domain of ICA69 Interacts with PICK1 and Acts on Trafficking of PICK1-PKC? Complex and Cerebellar Plasticity
    Article Snippet: Amylose resin (New England Biolabs, Hitchin, UK) was added to cleared lysates. .. Purified MBP fusion proteins were then dialyzed against internal solution.

    Article Title: The two TRAPP complexes of metazoans have distinct roles and act on different Rab GTPases
    Article Snippet: The TRAPPC9 (aa23–514)-6×His-MBP fusion protein was purified from the cleared lysate with amylose resin (E8021S; New England Biolabs, Inc.), and coupled with CNBr-activated Sepharose 4B beads. .. The TRAPPC9 (aa23–514)-6×His-MBP fusion protein was purified from the cleared lysate with amylose resin (E8021S; New England Biolabs, Inc.), and coupled with CNBr-activated Sepharose 4B beads.

    Article Title: Structure Based Identification and Characterization of Flavonoids That Disrupt Human Papillomavirus-16 E6 Function
    Article Snippet: Cell lysate was cleared by centrifugation at 15000xg, 4°C for 30 min and equilibrated with amylose resin beads (New England Biolabs, Inc.) by inversion for 3 hrs at 4°C. .. Bound MBP-E6 proteins were eluted using a final concentration of 20 mM maltose in 5 bed volumes of wash buffer.

    Polymerase Chain Reaction:

    Article Title: Autophosphorylation of Ser-6 via an intermolecular mechanism is important for the rapid reduction of NtCDPK1 kinase activity for substrate RSG
    Article Snippet: PCR products were cloned into pGEX-4T-1 (GE Healthcare). .. Glutathione S -transferase (GST)-NtCDPK1s, maltose-binding protein (MBP)-RSG, and 10×His-tagged (His)-NtCDPK1 were expressed in Escherichia coli harbouring pGEX-4T-1-NtCDPK1, pMALc2-RSG, or pET16b-NtCDPK1, respectively, and purified by glutathione-Sepharose 4B (GE Healthcare), Amylose Resin (New England Biolabs), or COSMOGEL His-Accept (Nacalai Tesque), respectively.

    Article Title: DER containing two consecutive GTP-binding domains plays an essential role in chloroplast ribosomal RNA processing and ribosome biogenesis in higher plants
    Article Snippet: To purify recombinant proteins of NbDER and its mutants for GTPase assays, the corresponding NbDER cDNA fragments were PCR-amplified and cloned into the pMALTM c2 or pMAL-c2 vector (New England Biolabs). .. The MBP fusion proteins were purified using amylose resin following the manufacturer’s instructions (New England Biolabs).

    Article Title: Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases
    Article Snippet: Other materials were provided by the following suppliers: SMART RACE™ cDNA Amplification kit and Advantage 2 PCR System were from CLONTECH, Mountain View, California, USA. .. ; restriction enzymes were from Promega, Chilworth, Southampton, U.K.; chelating Sepharose fast flow resin and gel filtration columns were from GE Healthcare, Little Chalfont, Bucks, U.K.; pET14b was from Novagen, Madison, WI, USA; pMalC2 and amylose resin were from New England Biolabs, Beverly, Maine, USA: all RNA preservation systems were from AMBION, Austin, Texas, USA; RNA extraction kits were from QIAGEN, Crawley, West Sussex, UK; and primers were from Invitrogen, Paisley, UK.

    Sonication:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: The lysate was sonicated for three cycles of 30 s, setting 4, 50% duty cycle with a microtip (Vibracell, Sonics and Materials, Inc.) and subsequently centrifuged twice in an Oakridge tube at 9,000 × g for 30 min in a JA25.5 rotor. .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Article Title: C-Terminal Domain of ICA69 Interacts with PICK1 and Acts on Trafficking of PICK1-PKC? Complex and Cerebellar Plasticity
    Article Snippet: Lysates were liberated by sonication and solubilized by 2% Triton X-100 in STE buffer. .. Amylose resin (New England Biolabs, Hitchin, UK) was added to cleared lysates.

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: After the collection of cells by centrifugation (5,000 rpm for 5 min), the cells were resuspended and sonicated in low-salt column buffer (30 mM HEPES-KOH pH 7.4, 25 mM NaCl, 1 mM EDTA, 10 mM β-mercaptoethanol). .. To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C.

    Article Title: Structure Based Identification and Characterization of Flavonoids That Disrupt Human Papillomavirus-16 E6 Function
    Article Snippet: Cells were lysed in 50 mM Tris HCl pH 6.8, 300 mM NaCl, 0.1 % (v/v) NP-40, 2 mM DTT and 1x protease inhibitor cocktail (EDTA-free, Roche) and sonicated as described before. .. Cell lysate was cleared by centrifugation at 15000xg, 4°C for 30 min and equilibrated with amylose resin beads (New England Biolabs, Inc.) by inversion for 3 hrs at 4°C.

    Article Title: Novel interaction of Rab13 and Rab8 with endospanins
    Article Snippet: Cells were suspended in binding buffer (50 mM Tris, 300 mM NaCl, 5 mM MgCl2 , 1 mM EDTA, 10 mM β-mercaptoethanol, 0.1% Triton X-100 and protease inhibitors (Roche), pH 7.2) and lysed by sonication. .. Equimolar amounts of MBP-bait proteins (∼10 μg) were incubated with GST-target proteins (∼2 μg) for 2 h at 4 °C together with amylose resin beads (New England BioLabs).

    Recombinant:

    Article Title: Autophosphorylation of Ser-6 via an intermolecular mechanism is important for the rapid reduction of NtCDPK1 kinase activity for substrate RSG
    Article Snippet: Paragraph title: Preparation of recombinant proteins ... Glutathione S -transferase (GST)-NtCDPK1s, maltose-binding protein (MBP)-RSG, and 10×His-tagged (His)-NtCDPK1 were expressed in Escherichia coli harbouring pGEX-4T-1-NtCDPK1, pMALc2-RSG, or pET16b-NtCDPK1, respectively, and purified by glutathione-Sepharose 4B (GE Healthcare), Amylose Resin (New England Biolabs), or COSMOGEL His-Accept (Nacalai Tesque), respectively.

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: The protein concentration was determined using a Nanodrop 1000 (Thermo Scientific) and the concentrated protein was stored at 4 °C for up to 2 weeks without a noticeable decrease in activity. .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation. .. The resin was poured into a 1.5 × 12-cm chromatography column (Bio-Rad), washed with ∼100 column volumes of MBP buffer (20 mm Tris-HCl, pH 7.4, 0.2 m NaCl), and bound protein was eluted with 10 ml of MBP buffer containing 50 mm maltose.

    Article Title: Rac function is crucial for cell migration but is not required for spreading and focal adhesion formation
    Article Snippet: The plasmids used in this study are listed in supplementary material Table S1 . .. All recombinant proteins were overexpressed in E. coli BL21(DE3)pLysS (GE Healthcare) according to standard procedures, using glutathione–Sepharose (GE Healthcare) and amylose resin (NEB). .. Proteins bound to the matrix were snap frozen in 50 mM Tris-HCl, 150 mM NaCl, 5 mM MgCl2 , 1 mM DTT, protease inhibitors (Mini Complete, EDTA free, Roche), pH 7.5 supplemented with 10% glycerol.

    Article Title: Role of Tomato Lipoxygenase D in Wound-Induced Jasmonate Biosynthesis and Plant Immunity to Insect Herbivores
    Article Snippet: For plasmid construction of maltose binding protein (MBP) fusions with SlMYC2, the cDNA was amplified and cloned into the pMAL-c2 vector (New England Biolabs, Beverly, MA) via Bam HI and Pst I restriction sites. .. The MBP-SlMYC2 recombinant protein was expressed in the BL21 Escheichia coli (E. coli ) strain and purified by binding onto an amylose resin (New England Biolabs) column, according to the instructions provided by the manufacturer. .. The 50-bp TomLoxD promoter probes containing G-box-like motif at the -369 site were synthesized and labeled with biotin at the 3′ end (Invitrogen), which containing the same sequences as that of the competitor probes without biotin-labled, while the mutated labeled probes were deleted the G-box-like motif.

    Article Title: Mitotic spindle scaling during Xenopus development by kif2a and importin ?
    Article Snippet: The NLS mutant had three charge switch mutations, K427E, K428E, and K430E, introduced into the NLS sequence identified by Quikchange mutagenesis (Stratagene/Agilent Technologies, Santa Clara CA, USA). .. 1 µM recombinant kif2a or kif2a ΔNLS as an MBP fusion protein or MBP alone was added to extracts and incubated for 15 min. 75 µl of Amylose resin (New England Biolabs) was then incubated for 15 min to retrieved MBP or fusion proteins. .. Resin was washed with a total of 25 bed volumes of phosphate-buffered saline with 0.01% Tween and eluted in SDS-PAGE loading buffer.

    Article Title: DER containing two consecutive GTP-binding domains plays an essential role in chloroplast ribosomal RNA processing and ribosome biogenesis in higher plants
    Article Snippet: Paragraph title: Purification of recombinant proteins ... The MBP fusion proteins were purified using amylose resin following the manufacturer’s instructions (New England Biolabs).

    Article Title: The Self-Interaction of a Nodavirus Replicase Is Enhanced by Mitochondrial Membrane Lipids
    Article Snippet: Briefly, to obtain soluble recombinant protein, Maltose-binding protein (MBP)-tagged full-length protein A and its mutants as well as the negative control protein MBP were expressed in Escherichia coli strain TB1 at 20°C in the presence of 0.2 mM IPTG. .. The proteins in the supernatant were purified using amylose resin (New England BioLabs) according to the manufacturer’s protocol and concentrated using Amicon Ultra-15 filters (Millipore, Schwalbach, Germany), and the buffer was exchanged to the hypotonic buffer (1 mM HEPES [pH 7.4], 0.1 mM EDTA, 15 mM NaCl, 1 mM DTT).

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: Paragraph title: Recombinant protein purification from E. coli and in vitro pull-down assay ... To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C.

    Article Title: Structure of CbpA J-Domain Bound to the Regulatory Protein CbpM Explains Its Specificity and Suggests Evolutionary Link between CbpM and Transcriptional Regulators
    Article Snippet: Recombinant CbpA, CbpM and CbpAJdom were purified by standard immobilized metal affinity chromatography using Ni2+ -NTA resin (Qiagen) using 50 mM TRIS-HCl, pH 8.0 with 250–300 mM NaCl and eluted with the same buffer containing 200–350 mM imidazole. .. For CbpM and CbpA, the protein samples were also passed through amylose resin (New England Biolabs Inc).

    Article Title: 18S rRNA processing requires base pairings of snR30 H/ACA snoRNA to eukaryote-specific 18S sequences
    Article Snippet: The spheroplast suspension was transferred onto ice, supplemented with 100 μg/ml AMT psoralen (4′-aminomethyl-4,5′,8-trimethylpsoralen) (HRI Associates Inc.), incubated in the dark for 15 min and irradiated with UV light (365 nm, ∼25 mW/cm2 ) for 15 min. Total RNA was extracted from the irradiated spheroplasts. .. For affinity selection of M2-R30 RNA, about 20 μg of purified recombinant MS2-MBP protein was bound to 20 μl of amylose resin (New England BioLabs) in NET2 buffer (200 mM NaCl, 0.05% Nonidet P-40 and 20 mM Tris–HCl pH 7.4) as described ( ). .. The beads were washed three times with 500 μl of NET2 and about 100 μg of cellular RNA was added in 50 μl of NET2.

    In Vivo:

    Article Title: 18S rRNA processing requires base pairings of snR30 H/ACA snoRNA to eukaryote-specific 18S sequences
    Article Snippet: Paragraph title: In vivo crosslinking ... For affinity selection of M2-R30 RNA, about 20 μg of purified recombinant MS2-MBP protein was bound to 20 μl of amylose resin (New England BioLabs) in NET2 buffer (200 mM NaCl, 0.05% Nonidet P-40 and 20 mM Tris–HCl pH 7.4) as described ( ).

    Pull Down Assay:

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: Paragraph title: Recombinant protein purification from E. coli and in vitro pull-down assay ... To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C.

    Mutagenesis:

    Article Title: Mechanistic insights into the role of Hop2-Mnd1 in meiotic homologous DNA pairing
    Article Snippet: Mouse Hop2, Hop2–Mnd1 and their mutant derivatives were expressed in Escherichia coli and purified as described ( , ). .. Singly and doubly MBP-tagged Hop2–Mnd1 species were expressed in E. coli and purified as for Hop2–Mnd1 but with an additional affinity step using Amylose resin [New England Biolabs; for Hop2-(MBP–Mnd1)] or anti-Flag resin [Sigma-Aldrich; for (MBP–Hop2)-Mnd1 and (MBP–Hop2)-(MBP–Mnd1)] following the protocols provided by the manufacturers.

    Negative Control:

    Article Title: The Self-Interaction of a Nodavirus Replicase Is Enhanced by Mitochondrial Membrane Lipids
    Article Snippet: Briefly, to obtain soluble recombinant protein, Maltose-binding protein (MBP)-tagged full-length protein A and its mutants as well as the negative control protein MBP were expressed in Escherichia coli strain TB1 at 20°C in the presence of 0.2 mM IPTG. .. The proteins in the supernatant were purified using amylose resin (New England BioLabs) according to the manufacturer’s protocol and concentrated using Amicon Ultra-15 filters (Millipore, Schwalbach, Germany), and the buffer was exchanged to the hypotonic buffer (1 mM HEPES [pH 7.4], 0.1 mM EDTA, 15 mM NaCl, 1 mM DTT).

    Electrophoretic Mobility Shift Assay:

    Article Title: Role of Tomato Lipoxygenase D in Wound-Induced Jasmonate Biosynthesis and Plant Immunity to Insect Herbivores
    Article Snippet: Paragraph title: Electrophoretic Mobility Shift Assay ... The MBP-SlMYC2 recombinant protein was expressed in the BL21 Escheichia coli (E. coli ) strain and purified by binding onto an amylose resin (New England Biolabs) column, according to the instructions provided by the manufacturer.

    Titration:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: For the time course and titration experiments, the Ni2+ -NTA eluate was polished by gel filtration chromatography on a HiLoad 26/60 Superdex 200 prep-grade column equilibrated in standard assay buffer (10 mm HEPES-Na+ , 1 mm EDTA-Na+ , 50 mm NaCl, pH 7.4). .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Protein Purification:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: Paragraph title: Protein Purification ... To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Article Title: Mechanistic insights into the role of Hop2-Mnd1 in meiotic homologous DNA pairing
    Article Snippet: Paragraph title: Protein purification procedures ... Singly and doubly MBP-tagged Hop2–Mnd1 species were expressed in E. coli and purified as for Hop2–Mnd1 but with an additional affinity step using Amylose resin [New England Biolabs; for Hop2-(MBP–Mnd1)] or anti-Flag resin [Sigma-Aldrich; for (MBP–Hop2)-Mnd1 and (MBP–Hop2)-(MBP–Mnd1)] following the protocols provided by the manufacturers.

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: Paragraph title: Recombinant protein purification from E. coli and in vitro pull-down assay ... To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C.

    Sequencing:

    Article Title: The two TRAPP complexes of metazoans have distinct roles and act on different Rab GTPases
    Article Snippet: To purify TRAPPC9 antisera, the N terminus of TRAPPC9 corresponding with amino acids 23–514 of GenBank accession number was fused at its N terminus to His-MBP by cloning the corresponding region of the coding sequence of cDNA clone RE66325 into the expression vector pETM-41. .. The TRAPPC9 (aa23–514)-6×His-MBP fusion protein was purified from the cleared lysate with amylose resin (E8021S; New England Biolabs, Inc.), and coupled with CNBr-activated Sepharose 4B beads.

    Article Title: Identification of multiple binding sites for the THAP domain of the Galileo transposase in the long terminal inverted-repeats
    Article Snippet: Since no ancestral sequence was reconstructed for the D. buzzatii domain, the 450 bp THAP sequence (150 amino acid) was cloned directly in the pOPINM expression vector. .. The supernatant was loaded onto an amylose resin column (New England Biolabs).

    Crystallization Assay:

    Article Title: Structure of CbpA J-Domain Bound to the Regulatory Protein CbpM Explains Its Specificity and Suggests Evolutionary Link between CbpM and Transcriptional Regulators
    Article Snippet: For CbpM and CbpA, the protein samples were also passed through amylose resin (New England Biolabs Inc). .. For CbpM and CbpA, the protein samples were also passed through amylose resin (New England Biolabs Inc).

    Affinity Chromatography:

    Article Title: Structure of CbpA J-Domain Bound to the Regulatory Protein CbpM Explains Its Specificity and Suggests Evolutionary Link between CbpM and Transcriptional Regulators
    Article Snippet: Recombinant CbpA, CbpM and CbpAJdom were purified by standard immobilized metal affinity chromatography using Ni2+ -NTA resin (Qiagen) using 50 mM TRIS-HCl, pH 8.0 with 250–300 mM NaCl and eluted with the same buffer containing 200–350 mM imidazole. .. For CbpM and CbpA, the protein samples were also passed through amylose resin (New England Biolabs Inc).

    Staining:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: Peak protein fractions were identified by SDS-PAGE and Coomassie staining, and concentrated using a 30-kDa molecular mass cut-off Vivaspin 15 concentrator (Sartorium Stedim Biotech). .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Article Title: A Cell Cycle and Nutritional Checkpoint Controlling Bacterial Surface Adhesion
    Article Snippet: The supernatant was then mixed with 500 µl of Amylose resin (New England Biolabs) pre-equilibrated with Tris-NaCl buffer, which allowed for binding of MBP domains. .. Two stringent washing steps were performed with Tris-NaCl buffer containing 10 mM or 75 mM imidazole followed by elution with 100 µl of 1 M imidazole Tris-NaCL buffer.

    Purification:

    Article Title: Autophosphorylation of Ser-6 via an intermolecular mechanism is important for the rapid reduction of NtCDPK1 kinase activity for substrate RSG
    Article Snippet: PCR products were cloned into pGEX-4T-1 (GE Healthcare). .. Glutathione S -transferase (GST)-NtCDPK1s, maltose-binding protein (MBP)-RSG, and 10×His-tagged (His)-NtCDPK1 were expressed in Escherichia coli harbouring pGEX-4T-1-NtCDPK1, pMALc2-RSG, or pET16b-NtCDPK1, respectively, and purified by glutathione-Sepharose 4B (GE Healthcare), Amylose Resin (New England Biolabs), or COSMOGEL His-Accept (Nacalai Tesque), respectively. .. GST-NtCDPK1 proteins (2.0 μg) were reacted in autophosphorylation buffer containing 20 mM Tris-HCl, pH 7.5, 10 mM MgCl2 , 0.5 mM CaCl2 , 0.1% (v/v) Triton X-100, 0.05% (v/v) β-mercaptoethanol, and 1 mM ATP at 30°C for 1 h. Catalytically inactive forms of GST-NtCDPK1, GST-NtCDPK1 D219N, GST-NtCDPK1 S6A/D219N, GST-NtCDPK1 T21A/D219N and GST-NtCDPK1 S6A/T21A/D219N, were autophosphorylated by His-NtCDPK1 in autophosphorylation buffer at 30°C for 1 h. The phosphorylation state was examined using Phosphate-binding tag (Phos-tag) SDS-PAGE [ ].

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation. .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Article Title: C-Terminal Domain of ICA69 Interacts with PICK1 and Acts on Trafficking of PICK1-PKC? Complex and Cerebellar Plasticity
    Article Snippet: Paragraph title: Purification of MBP fusion protein ... Amylose resin (New England Biolabs, Hitchin, UK) was added to cleared lysates.

    Article Title: Role of Tomato Lipoxygenase D in Wound-Induced Jasmonate Biosynthesis and Plant Immunity to Insect Herbivores
    Article Snippet: For plasmid construction of maltose binding protein (MBP) fusions with SlMYC2, the cDNA was amplified and cloned into the pMAL-c2 vector (New England Biolabs, Beverly, MA) via Bam HI and Pst I restriction sites. .. The MBP-SlMYC2 recombinant protein was expressed in the BL21 Escheichia coli (E. coli ) strain and purified by binding onto an amylose resin (New England Biolabs) column, according to the instructions provided by the manufacturer. .. The 50-bp TomLoxD promoter probes containing G-box-like motif at the -369 site were synthesized and labeled with biotin at the 3′ end (Invitrogen), which containing the same sequences as that of the competitor probes without biotin-labled, while the mutated labeled probes were deleted the G-box-like motif.

    Article Title: WRB and CAML Are Necessary and Sufficient to Mediate Tail-Anchored Protein Targeting to the ER Membrane
    Article Snippet: Paragraph title: Protein Expression and Purification ... The supernatant was loaded onto an amylose resin (NEB) column.

    Article Title: DER containing two consecutive GTP-binding domains plays an essential role in chloroplast ribosomal RNA processing and ribosome biogenesis in higher plants
    Article Snippet: To purify recombinant proteins of NbDER and its mutants for GTPase assays, the corresponding NbDER cDNA fragments were PCR-amplified and cloned into the pMALTM c2 or pMAL-c2 vector (New England Biolabs). .. The MBP fusion proteins were purified using amylose resin following the manufacturer’s instructions (New England Biolabs). .. Purified proteins of MBP:NbDER, MBP:PM1, MBP:PM2, and MBP:PM1/2 were concentrated using Amicon Ultra Centrifugal Filters (Millipore).

    Article Title: Mechanistic insights into the role of Hop2-Mnd1 in meiotic homologous DNA pairing
    Article Snippet: Hop2 1 −84 , purified without the hydroxyapatite step, elutes from SP Sepharose at 100–200 mM KCl and from Mono S at 120–160 mM KCl. .. Singly and doubly MBP-tagged Hop2–Mnd1 species were expressed in E. coli and purified as for Hop2–Mnd1 but with an additional affinity step using Amylose resin [New England Biolabs; for Hop2-(MBP–Mnd1)] or anti-Flag resin [Sigma-Aldrich; for (MBP–Hop2)-Mnd1 and (MBP–Hop2)-(MBP–Mnd1)] following the protocols provided by the manufacturers. .. Human DMC1 was purified from insect cells infected with a recombinant DMC1 baculovirus, as described ( ).

    Article Title: The Self-Interaction of a Nodavirus Replicase Is Enhanced by Mitochondrial Membrane Lipids
    Article Snippet: Cell pellets were resuspended in binding buffer (20 mM Tris-HCl [pH 7.4], 200 mM NaCl, 1 mM EDTA, 10 mM 2-Mercaptoethanol) supplemented with 1.5% Triton-X 100 and protease inhibitors cocktail (Sigma, St. Louis, Mo, USA). .. The proteins in the supernatant were purified using amylose resin (New England BioLabs) according to the manufacturer’s protocol and concentrated using Amicon Ultra-15 filters (Millipore, Schwalbach, Germany), and the buffer was exchanged to the hypotonic buffer (1 mM HEPES [pH 7.4], 0.1 mM EDTA, 15 mM NaCl, 1 mM DTT). .. For in vitro translation, His-tagged proteins were translated using nuclease-treated rabbit reticulocyte lysates (Promega) according to the manufacturer’s protocol.

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: Recombinant MBP-tagged helicase proteins, the MBP-tagged TBSV p33 and p92 replication proteins and several truncated MBP-tagged p33C derivatives (described earlier) were expressed in E. coli and purified as published earlier with modifications , . .. To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C.

    Article Title: Structure of CbpA J-Domain Bound to the Regulatory Protein CbpM Explains Its Specificity and Suggests Evolutionary Link between CbpM and Transcriptional Regulators
    Article Snippet: Paragraph title: Protein Expression and Purification ... For CbpM and CbpA, the protein samples were also passed through amylose resin (New England Biolabs Inc).

    Article Title: Cep63 and Cep152 Cooperate to Ensure Centriole Duplication
    Article Snippet: Resin was washed with PBS, and proteins were eluted by boiling in Laemmli buffer (Bio-Rad). .. MBP and MBP-Cep63 were expressed in BL21 CodonPlus (DE3) RIL E. coli (Stratagene) at 16°C for 3 hours after induction with 0.3 mM IPTG (Isopropyl β-D-1-thiogalactopyranoside), and purified using amylose resin according the manufacturer’s protocol (NEB). .. Cells were grown on glass coverslips; fixed in −20°C methanol for at least 10 minutes; rehydrated in PBS 0.01% Triton X-100 (TX); and permeabilised with PBS 0.2% TX.

    Article Title: 18S rRNA processing requires base pairings of snR30 H/ACA snoRNA to eukaryote-specific 18S sequences
    Article Snippet: The spheroplast suspension was transferred onto ice, supplemented with 100 μg/ml AMT psoralen (4′-aminomethyl-4,5′,8-trimethylpsoralen) (HRI Associates Inc.), incubated in the dark for 15 min and irradiated with UV light (365 nm, ∼25 mW/cm2 ) for 15 min. Total RNA was extracted from the irradiated spheroplasts. .. For affinity selection of M2-R30 RNA, about 20 μg of purified recombinant MS2-MBP protein was bound to 20 μl of amylose resin (New England BioLabs) in NET2 buffer (200 mM NaCl, 0.05% Nonidet P-40 and 20 mM Tris–HCl pH 7.4) as described ( ). .. The beads were washed three times with 500 μl of NET2 and about 100 μg of cellular RNA was added in 50 μl of NET2.

    Article Title: The two TRAPP complexes of metazoans have distinct roles and act on different Rab GTPases
    Article Snippet: Cells were harvested and resuspended in column buffer (20 mM Tris-HCl, pH 7.4, 200 mM NaCl, 0.1 mM EDTA, 10 mM DTT, and cOmplete protease inhibitors). .. The TRAPPC9 (aa23–514)-6×His-MBP fusion protein was purified from the cleared lysate with amylose resin (E8021S; New England Biolabs, Inc.), and coupled with CNBr-activated Sepharose 4B beads. .. TRAPPC9 antibodies were bound to the beads and then eluted with 0.1 M glycine, pH 2.5, and fractions were neutralized with 1 M Tris, pH 8.8.

    SDS Page:

    Article Title: Identification of a Plasmodium falciparu
    Article Snippet: Peak protein fractions were identified by SDS-PAGE and Coomassie staining, and concentrated using a 30-kDa molecular mass cut-off Vivaspin 15 concentrator (Sartorium Stedim Biotech). .. To purify recombinant proteins used for the remaining transfer assays, the Ni2+ -NTA eluate was added to ∼1 ml of amylose resin (New England Biolabs) and incubated at 4 °C overnight with agitation.

    Article Title: C-Terminal Domain of ICA69 Interacts with PICK1 and Acts on Trafficking of PICK1-PKC? Complex and Cerebellar Plasticity
    Article Snippet: Amylose resin (New England Biolabs, Hitchin, UK) was added to cleared lysates. .. Fusion protein concentration was determined using a BCA kit assay (Thermo).

    Article Title: A Cell Cycle and Nutritional Checkpoint Controlling Bacterial Surface Adhesion
    Article Snippet: The supernatant was then mixed with 500 µl of Amylose resin (New England Biolabs) pre-equilibrated with Tris-NaCl buffer, which allowed for binding of MBP domains. .. Two stringent washing steps were performed with Tris-NaCl buffer containing 10 mM or 75 mM imidazole followed by elution with 100 µl of 1 M imidazole Tris-NaCL buffer.

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C. .. To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C.

    Article Title: Cep63 and Cep152 Cooperate to Ensure Centriole Duplication
    Article Snippet: Resin was washed with PBS; proteins were eluted from the resin by boiling in Laemmli buffer (Bio-Rad) then analysed by SDS-PAGE and Western blotting. .. MBP and MBP-Cep63 were expressed in BL21 CodonPlus (DE3) RIL E. coli (Stratagene) at 16°C for 3 hours after induction with 0.3 mM IPTG (Isopropyl β-D-1-thiogalactopyranoside), and purified using amylose resin according the manufacturer’s protocol (NEB).

    Plasmid Preparation:

    Article Title: Autophosphorylation of Ser-6 via an intermolecular mechanism is important for the rapid reduction of NtCDPK1 kinase activity for substrate RSG
    Article Snippet: NtCDPK1 S6A/D219N and T21A/D219N were generated using overlap extension PCR and seamless ligation cloning extract (SLiCE) [ ] with a plasmid harbouring full-length NtCDPK1 as a template and appropriate primers ( ). .. Glutathione S -transferase (GST)-NtCDPK1s, maltose-binding protein (MBP)-RSG, and 10×His-tagged (His)-NtCDPK1 were expressed in Escherichia coli harbouring pGEX-4T-1-NtCDPK1, pMALc2-RSG, or pET16b-NtCDPK1, respectively, and purified by glutathione-Sepharose 4B (GE Healthcare), Amylose Resin (New England Biolabs), or COSMOGEL His-Accept (Nacalai Tesque), respectively.

    Article Title: Role of Tomato Lipoxygenase D in Wound-Induced Jasmonate Biosynthesis and Plant Immunity to Insect Herbivores
    Article Snippet: For plasmid construction of maltose binding protein (MBP) fusions with SlMYC2, the cDNA was amplified and cloned into the pMAL-c2 vector (New England Biolabs, Beverly, MA) via Bam HI and Pst I restriction sites. .. The MBP-SlMYC2 recombinant protein was expressed in the BL21 Escheichia coli (E. coli ) strain and purified by binding onto an amylose resin (New England Biolabs) column, according to the instructions provided by the manufacturer.

    Article Title: DER containing two consecutive GTP-binding domains plays an essential role in chloroplast ribosomal RNA processing and ribosome biogenesis in higher plants
    Article Snippet: To purify recombinant proteins of NbDER and its mutants for GTPase assays, the corresponding NbDER cDNA fragments were PCR-amplified and cloned into the pMALTM c2 or pMAL-c2 vector (New England Biolabs). .. The MBP fusion proteins were purified using amylose resin following the manufacturer’s instructions (New England Biolabs).

    Article Title: 18S rRNA processing requires base pairings of snR30 H/ACA snoRNA to eukaryote-specific 18S sequences
    Article Snippet: Yeast GAL∷snR30 cells carrying the pMS2-R30 expression plasmid were grown in YPD liquid medium containing 2% sucrose at 30°C until OD600 0.8. .. For affinity selection of M2-R30 RNA, about 20 μg of purified recombinant MS2-MBP protein was bound to 20 μl of amylose resin (New England BioLabs) in NET2 buffer (200 mM NaCl, 0.05% Nonidet P-40 and 20 mM Tris–HCl pH 7.4) as described ( ).

    Article Title: The two TRAPP complexes of metazoans have distinct roles and act on different Rab GTPases
    Article Snippet: To purify TRAPPC9 antisera, the N terminus of TRAPPC9 corresponding with amino acids 23–514 of GenBank accession number was fused at its N terminus to His-MBP by cloning the corresponding region of the coding sequence of cDNA clone RE66325 into the expression vector pETM-41. .. The TRAPPC9 (aa23–514)-6×His-MBP fusion protein was purified from the cleared lysate with amylose resin (E8021S; New England Biolabs, Inc.), and coupled with CNBr-activated Sepharose 4B beads.

    Article Title: Identification of multiple binding sites for the THAP domain of the Galileo transposase in the long terminal inverted-repeats
    Article Snippet: Since no ancestral sequence was reconstructed for the D. buzzatii domain, the 450 bp THAP sequence (150 amino acid) was cloned directly in the pOPINM expression vector. .. The supernatant was loaded onto an amylose resin column (New England Biolabs).

    Irradiation:

    Article Title: 18S rRNA processing requires base pairings of snR30 H/ACA snoRNA to eukaryote-specific 18S sequences
    Article Snippet: The spheroplast suspension was transferred onto ice, supplemented with 100 μg/ml AMT psoralen (4′-aminomethyl-4,5′,8-trimethylpsoralen) (HRI Associates Inc.), incubated in the dark for 15 min and irradiated with UV light (365 nm, ∼25 mW/cm2 ) for 15 min. Total RNA was extracted from the irradiated spheroplasts. .. For affinity selection of M2-R30 RNA, about 20 μg of purified recombinant MS2-MBP protein was bound to 20 μl of amylose resin (New England BioLabs) in NET2 buffer (200 mM NaCl, 0.05% Nonidet P-40 and 20 mM Tris–HCl pH 7.4) as described ( ).

    RNA Extraction:

    Article Title: Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases
    Article Snippet: Other materials were provided by the following suppliers: SMART RACE™ cDNA Amplification kit and Advantage 2 PCR System were from CLONTECH, Mountain View, California, USA. .. ; restriction enzymes were from Promega, Chilworth, Southampton, U.K.; chelating Sepharose fast flow resin and gel filtration columns were from GE Healthcare, Little Chalfont, Bucks, U.K.; pET14b was from Novagen, Madison, WI, USA; pMalC2 and amylose resin were from New England Biolabs, Beverly, Maine, USA: all RNA preservation systems were from AMBION, Austin, Texas, USA; RNA extraction kits were from QIAGEN, Crawley, West Sussex, UK; and primers were from Invitrogen, Paisley, UK. .. Coryphaenoides armatus (rat tail) specimens were caught by otter trawl in the Atlantic from the Porcupine Seabight from a depth of 3986 m – 4016 m, September 2000, on RRS Discovery Cruise 250.

    Copurification:

    Article Title: A Cell Cycle and Nutritional Checkpoint Controlling Bacterial Surface Adhesion
    Article Snippet: Paragraph title: Protein co-expression and co-purification ... The supernatant was then mixed with 500 µl of Amylose resin (New England Biolabs) pre-equilibrated with Tris-NaCl buffer, which allowed for binding of MBP domains.

    In Vitro:

    Article Title: The Expanding Functions of Cellular Helicases: The Tombusvirus RNA Replication Enhancer Co-opts the Plant eIF4AIII-Like AtRH2 and the DDX5-Like AtRH5 DEAD-Box RNA Helicases to Promote Viral Asymmetric RNA Replication
    Article Snippet: Paragraph title: Recombinant protein purification from E. coli and in vitro pull-down assay ... To remove cells debris, the lysate was centrifuged at 14,000 rpm for 5 min, followed by supernatant incubation with amylose resin (NEB) for 15 min at 4°C.

    Article Title: Novel interaction of Rab13 and Rab8 with endospanins
    Article Snippet: Paragraph title: In vitro pull-down experiments ... Equimolar amounts of MBP-bait proteins (∼10 μg) were incubated with GST-target proteins (∼2 μg) for 2 h at 4 °C together with amylose resin beads (New England BioLabs).

    Electrophoresis:

    Article Title: C-Terminal Domain of ICA69 Interacts with PICK1 and Acts on Trafficking of PICK1-PKC? Complex and Cerebellar Plasticity
    Article Snippet: Amylose resin (New England Biolabs, Hitchin, UK) was added to cleared lysates. .. Fusion protein concentration was determined using a BCA kit assay (Thermo).

    Article Title: A Cell Cycle and Nutritional Checkpoint Controlling Bacterial Surface Adhesion
    Article Snippet: The supernatant was then mixed with 500 µl of Amylose resin (New England Biolabs) pre-equilibrated with Tris-NaCl buffer, which allowed for binding of MBP domains. .. Two stringent washing steps were performed with Tris-NaCl buffer containing 10 mM or 75 mM imidazole followed by elution with 100 µl of 1 M imidazole Tris-NaCL buffer.

    Selection:

    Article Title: 18S rRNA processing requires base pairings of snR30 H/ACA snoRNA to eukaryote-specific 18S sequences
    Article Snippet: The spheroplast suspension was transferred onto ice, supplemented with 100 μg/ml AMT psoralen (4′-aminomethyl-4,5′,8-trimethylpsoralen) (HRI Associates Inc.), incubated in the dark for 15 min and irradiated with UV light (365 nm, ∼25 mW/cm2 ) for 15 min. Total RNA was extracted from the irradiated spheroplasts. .. For affinity selection of M2-R30 RNA, about 20 μg of purified recombinant MS2-MBP protein was bound to 20 μl of amylose resin (New England BioLabs) in NET2 buffer (200 mM NaCl, 0.05% Nonidet P-40 and 20 mM Tris–HCl pH 7.4) as described ( ). .. The beads were washed three times with 500 μl of NET2 and about 100 μg of cellular RNA was added in 50 μl of NET2.

    Ethanol Precipitation:

    Article Title: 18S rRNA processing requires base pairings of snR30 H/ACA snoRNA to eukaryote-specific 18S sequences
    Article Snippet: For affinity selection of M2-R30 RNA, about 20 μg of purified recombinant MS2-MBP protein was bound to 20 μl of amylose resin (New England BioLabs) in NET2 buffer (200 mM NaCl, 0.05% Nonidet P-40 and 20 mM Tris–HCl pH 7.4) as described ( ). .. The beads were incubated on ice for 1 h, washed twice with 500 μl of cold low-salt buffer (10 mM NaCl, 0.5 mM EDTA, 2.0 mM DTT and 20 mM Tris–HCl pH 7.4).

    Preserving:

    Article Title: Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases
    Article Snippet: Other materials were provided by the following suppliers: SMART RACE™ cDNA Amplification kit and Advantage 2 PCR System were from CLONTECH, Mountain View, California, USA. .. ; restriction enzymes were from Promega, Chilworth, Southampton, U.K.; chelating Sepharose fast flow resin and gel filtration columns were from GE Healthcare, Little Chalfont, Bucks, U.K.; pET14b was from Novagen, Madison, WI, USA; pMalC2 and amylose resin were from New England Biolabs, Beverly, Maine, USA: all RNA preservation systems were from AMBION, Austin, Texas, USA; RNA extraction kits were from QIAGEN, Crawley, West Sussex, UK; and primers were from Invitrogen, Paisley, UK. .. Coryphaenoides armatus (rat tail) specimens were caught by otter trawl in the Atlantic from the Porcupine Seabight from a depth of 3986 m – 4016 m, September 2000, on RRS Discovery Cruise 250.

    Spectrophotometry:

    Article Title: The Self-Interaction of a Nodavirus Replicase Is Enhanced by Mitochondrial Membrane Lipids
    Article Snippet: The proteins in the supernatant were purified using amylose resin (New England BioLabs) according to the manufacturer’s protocol and concentrated using Amicon Ultra-15 filters (Millipore, Schwalbach, Germany), and the buffer was exchanged to the hypotonic buffer (1 mM HEPES [pH 7.4], 0.1 mM EDTA, 15 mM NaCl, 1 mM DTT). .. For in vitro translation, His-tagged proteins were translated using nuclease-treated rabbit reticulocyte lysates (Promega) according to the manufacturer’s protocol.

    Produced:

    Article Title: Novel interaction of Rab13 and Rab8 with endospanins
    Article Snippet: 2.5 MBP- or GST-fusion proteins were produced by bacterial expression and cellular pellets were collected. .. Equimolar amounts of MBP-bait proteins (∼10 μg) were incubated with GST-target proteins (∼2 μg) for 2 h at 4 °C together with amylose resin beads (New England BioLabs).

    Concentration Assay:

    Article Title: Structure Based Identification and Characterization of Flavonoids That Disrupt Human Papillomavirus-16 E6 Function
    Article Snippet: Cell lysate was cleared by centrifugation at 15000xg, 4°C for 30 min and equilibrated with amylose resin beads (New England Biolabs, Inc.) by inversion for 3 hrs at 4°C. .. Bound beads were washed one time in lysis buffer and four times with wash buffer (50 mM Tris-HCl pH 6.8, 400 mM NaCl, 2 mM DTT, 1x protease inhibitor cocktail).

    Lysis:

    Article Title: WRB and CAML Are Necessary and Sufficient to Mediate Tail-Anchored Protein Targeting to the ER Membrane
    Article Snippet: After lysis with Avestin Emulsiflex-C5 aggregates were removed by centrifugation at 100,000 g , 30 minutes, 4°C. .. The supernatant was loaded onto an amylose resin (NEB) column.

    Article Title: Cep63 and Cep152 Cooperate to Ensure Centriole Duplication
    Article Snippet: After washing in column buffer, amylose resin was incubated with 1 mg cell lysate from cells 293 expressing GFP-Cep152 full length, N-, or C-terminal domains, in lysis buffer for 2 hours at 4°C. .. MBP and MBP-Cep63 were expressed in BL21 CodonPlus (DE3) RIL E. coli (Stratagene) at 16°C for 3 hours after induction with 0.3 mM IPTG (Isopropyl β-D-1-thiogalactopyranoside), and purified using amylose resin according the manufacturer’s protocol (NEB).

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    New England Biolabs amylose resin
    Amylose Resin, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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