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Millipore xmai
Xmai, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 4 article reviews
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xmai - by Bioz Stars, 2019-12
94/100 stars

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Clone Assay:

Article Title: Arabidopsis protein disulfide isomerase-8 is a type I endoplasmic reticulum transmembrane protein with thiol-disulfide oxidase activity
Article Snippet: SDS-PAGE and immunoblot detection of PDI8 was performed as described above, with each lane loaded with 20 μL of sample (~7.2 μg of microsomal protein). .. A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO). .. The PDI8 abb’ gene fragment was amplified from a full-length PDI8 cDNA using a forward primer with an XmaI site (5’-TGT CCC GGG AGA TGA TCA ATT CAC CCT CGA C-3’) and a reverse primer with a SalI site (5’-AAT GTC GAC CAT TGA GTT GAT AAA TCC CAT G-3’).

Article Title: Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine
Article Snippet: Despite this high degree of structural conservation, a marked specificity for the synthesis of A-PG or L-PG was observed for A-PGS and L-PGS in enzymatic assays using Ala-tRNAAla or Lys-tRNALys as substrates, respectively ( ). .. Base pairs 1627–2643 of ORF PA0920 from P . aeruginosa PAO1 and base pairs 1555–2550 of ORF yfiX from B . licheniformis DSM13 were PCR-amplified using oligonucleotide pairs 1 and 2 and 3 and 4 , respectively, and were cloned into the XmaI and SacI sites of pET52b(+) (Novagen) for expression with a cleavable N-terminal Strep -II-tag. .. The SerP (surface entropy reduction) server ( ) was used to identify mutations that may facilitate optimized crystallization of A-PGS (KGKE674 to AGAA674 ).

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The Quikchange II mutagenesis kit (Stratagene) was used to generate the various C-clamp mutants in the pAc-TCF/Pan expression vector. pAc-Arm* and the luciferase reporters, pGL3-nkd-intE and pGL3-6xHMG, have been described previously , . .. The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites. .. The pET vector encodes a C terminus 10xHis tag, which was used for protein purification.

Centrifugation:

Article Title: Arabidopsis protein disulfide isomerase-8 is a type I endoplasmic reticulum transmembrane protein with thiol-disulfide oxidase activity
Article Snippet: A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO). .. The pFLAG-PDI8abb’ construct and pFLAG-CTS empty vector were transformed into strain RI90.

Amplification:

Article Title: Rotavirus Replication Requires a Functional Proteasome for Effective Assembly of Viroplasms
Article Snippet: In order to construct the plasmid pVAX-T7-segment11-ribo-T7stop, indicated in the Results section as pVAX-segment 11, the pVAX vector (Invitrogen) was modified by deletion of the cytomegalovirus (CMV) promoter using NdeI and ApaI restriction enzymes and subsequent insertion of an oligonucleotide containing the XmaI, ApaI, and SacII restriction sites (5′-TATGGGTACCTACTCCCGGGTTACAGGGCCCGCGGAGGCC-3′; Sigma). .. In order to construct the plasmid pVAX-T7-segment11-ribo-T7stop, indicated in the Results section as pVAX-segment 11, the pVAX vector (Invitrogen) was modified by deletion of the cytomegalovirus (CMV) promoter using NdeI and ApaI restriction enzymes and subsequent insertion of an oligonucleotide containing the XmaI, ApaI, and SacII restriction sites (5′-TATGGGTACCTACTCCCGGGTTACAGGGCCCGCGGAGGCC-3′; Sigma).

Construct:

Article Title: Arabidopsis protein disulfide isomerase-8 is a type I endoplasmic reticulum transmembrane protein with thiol-disulfide oxidase activity
Article Snippet: SDS-PAGE and immunoblot detection of PDI8 was performed as described above, with each lane loaded with 20 μL of sample (~7.2 μg of microsomal protein). .. A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO). .. The PDI8 abb’ gene fragment was amplified from a full-length PDI8 cDNA using a forward primer with an XmaI site (5’-TGT CCC GGG AGA TGA TCA ATT CAC CCT CGA C-3’) and a reverse primer with a SalI site (5’-AAT GTC GAC CAT TGA GTT GAT AAA TCC CAT G-3’).

Article Title: Structural basis for ubiquitin-mediated antiviral signal activation by RIG-I
Article Snippet: Paragraph title: Plasmid constructs ... Wild type and variants of RIG-I 2CARD (residue 1–200) were subcloned between XmaI and HindIII restriction sites in pET47b (Novagen).

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The Quikchange II mutagenesis kit (Stratagene) was used to generate the various C-clamp mutants in the pAc-TCF/Pan expression vector. pAc-Arm* and the luciferase reporters, pGL3-nkd-intE and pGL3-6xHMG, have been described previously , . .. The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites. .. The pET vector encodes a C terminus 10xHis tag, which was used for protein purification.

Article Title: Rotavirus Replication Requires a Functional Proteasome for Effective Assembly of Viroplasms
Article Snippet: The NSP5 gene was derived from the OSU rotavirus strain, and the NSP2, VP1, VP2 and VP6 genes came from the SA11 strain. .. In order to construct the plasmid pVAX-T7-segment11-ribo-T7stop, indicated in the Results section as pVAX-segment 11, the pVAX vector (Invitrogen) was modified by deletion of the cytomegalovirus (CMV) promoter using NdeI and ApaI restriction enzymes and subsequent insertion of an oligonucleotide containing the XmaI, ApaI, and SacII restriction sites (5′-TATGGGTACCTACTCCCGGGTTACAGGGCCCGCGGAGGCC-3′; Sigma). .. Between the sites of XmaI and ApaI, a synthetic sequence (Genscript) has been inserted in order to clone the ribozyme and the T7 terminator (5′- ccCGGG TCGGCATGGCATCTCCACCTCCTCGCGGTCCGACCTGGGCATCCGAAGGAGGACGTCGTCCACTCGGATGGCTAAGGGAGAGCTcggatccGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATCGAGATCCtctaga gggccc -3′; capital letters indicate the ribozyme, and the T7 terminator and the XmaI and ApaI sites are underlined).

Luciferase:

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The Quikchange II mutagenesis kit (Stratagene) was used to generate the various C-clamp mutants in the pAc-TCF/Pan expression vector. pAc-Arm* and the luciferase reporters, pGL3-nkd-intE and pGL3-6xHMG, have been described previously , . .. The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites.

Activity Assay:

Article Title: Arabidopsis protein disulfide isomerase-8 is a type I endoplasmic reticulum transmembrane protein with thiol-disulfide oxidase activity
Article Snippet: A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO). .. The pFLAG-PDI8abb’ construct and pFLAG-CTS empty vector were transformed into strain RI90.

Expressing:

Article Title: Arabidopsis protein disulfide isomerase-8 is a type I endoplasmic reticulum transmembrane protein with thiol-disulfide oxidase activity
Article Snippet: SDS-PAGE and immunoblot detection of PDI8 was performed as described above, with each lane loaded with 20 μL of sample (~7.2 μg of microsomal protein). .. A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO). .. The PDI8 abb’ gene fragment was amplified from a full-length PDI8 cDNA using a forward primer with an XmaI site (5’-TGT CCC GGG AGA TGA TCA ATT CAC CCT CGA C-3’) and a reverse primer with a SalI site (5’-AAT GTC GAC CAT TGA GTT GAT AAA TCC CAT G-3’).

Article Title: Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine
Article Snippet: Despite this high degree of structural conservation, a marked specificity for the synthesis of A-PG or L-PG was observed for A-PGS and L-PGS in enzymatic assays using Ala-tRNAAla or Lys-tRNALys as substrates, respectively ( ). .. Base pairs 1627–2643 of ORF PA0920 from P . aeruginosa PAO1 and base pairs 1555–2550 of ORF yfiX from B . licheniformis DSM13 were PCR-amplified using oligonucleotide pairs 1 and 2 and 3 and 4 , respectively, and were cloned into the XmaI and SacI sites of pET52b(+) (Novagen) for expression with a cleavable N-terminal Strep -II-tag. .. The SerP (surface entropy reduction) server ( ) was used to identify mutations that may facilitate optimized crystallization of A-PGS (KGKE674 to AGAA674 ).

Article Title: Structural basis for ubiquitin-mediated antiviral signal activation by RIG-I
Article Snippet: Wild type and variants of RIG-I 2CARD (residue 1–200) were subcloned between XmaI and HindIII restriction sites in pET47b (Novagen). .. Wild type and variants of RIG-I 2CARD (residue 1–200) were subcloned between XmaI and HindIII restriction sites in pET47b (Novagen).

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The Quikchange II mutagenesis kit (Stratagene) was used to generate the various C-clamp mutants in the pAc-TCF/Pan expression vector. pAc-Arm* and the luciferase reporters, pGL3-nkd-intE and pGL3-6xHMG, have been described previously , . .. The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites. .. The pET vector encodes a C terminus 10xHis tag, which was used for protein purification.

ALP Activity Assay:

Article Title: Arabidopsis protein disulfide isomerase-8 is a type I endoplasmic reticulum transmembrane protein with thiol-disulfide oxidase activity
Article Snippet: Paragraph title: Alkaline phosphatase activity assay ... A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO).

Modification:

Article Title: Rotavirus Replication Requires a Functional Proteasome for Effective Assembly of Viroplasms
Article Snippet: The NSP5 gene was derived from the OSU rotavirus strain, and the NSP2, VP1, VP2 and VP6 genes came from the SA11 strain. .. In order to construct the plasmid pVAX-T7-segment11-ribo-T7stop, indicated in the Results section as pVAX-segment 11, the pVAX vector (Invitrogen) was modified by deletion of the cytomegalovirus (CMV) promoter using NdeI and ApaI restriction enzymes and subsequent insertion of an oligonucleotide containing the XmaI, ApaI, and SacII restriction sites (5′-TATGGGTACCTACTCCCGGGTTACAGGGCCCGCGGAGGCC-3′; Sigma). .. Between the sites of XmaI and ApaI, a synthetic sequence (Genscript) has been inserted in order to clone the ribozyme and the T7 terminator (5′- ccCGGG TCGGCATGGCATCTCCACCTCCTCGCGGTCCGACCTGGGCATCCGAAGGAGGACGTCGTCCACTCGGATGGCTAAGGGAGAGCTcggatccGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATCGAGATCCtctaga gggccc -3′; capital letters indicate the ribozyme, and the T7 terminator and the XmaI and ApaI sites are underlined).

Western Blot:

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites. .. For transgenic flies, the TCF/Pan ORF was cloned from the pAc-TCF/Pan constructs into the KpnI and XbaI restriction sites of the pUAST vector.

Transformation Assay:

Article Title: Arabidopsis protein disulfide isomerase-8 is a type I endoplasmic reticulum transmembrane protein with thiol-disulfide oxidase activity
Article Snippet: A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO). .. A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO).

Article Title: Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine
Article Snippet: Base pairs 1627–2643 of ORF PA0920 from P . aeruginosa PAO1 and base pairs 1555–2550 of ORF yfiX from B . licheniformis DSM13 were PCR-amplified using oligonucleotide pairs 1 and 2 and 3 and 4 , respectively, and were cloned into the XmaI and SacI sites of pET52b(+) (Novagen) for expression with a cleavable N-terminal Strep -II-tag. .. Base pairs 1627–2643 of ORF PA0920 from P . aeruginosa PAO1 and base pairs 1555–2550 of ORF yfiX from B . licheniformis DSM13 were PCR-amplified using oligonucleotide pairs 1 and 2 and 3 and 4 , respectively, and were cloned into the XmaI and SacI sites of pET52b(+) (Novagen) for expression with a cleavable N-terminal Strep -II-tag.

Derivative Assay:

Article Title: Rotavirus Replication Requires a Functional Proteasome for Effective Assembly of Viroplasms
Article Snippet: The NSP5 gene was derived from the OSU rotavirus strain, and the NSP2, VP1, VP2 and VP6 genes came from the SA11 strain. .. In order to construct the plasmid pVAX-T7-segment11-ribo-T7stop, indicated in the Results section as pVAX-segment 11, the pVAX vector (Invitrogen) was modified by deletion of the cytomegalovirus (CMV) promoter using NdeI and ApaI restriction enzymes and subsequent insertion of an oligonucleotide containing the XmaI, ApaI, and SacII restriction sites (5′-TATGGGTACCTACTCCCGGGTTACAGGGCCCGCGGAGGCC-3′; Sigma).

Sequencing:

Article Title: Arabidopsis protein disulfide isomerase-8 is a type I endoplasmic reticulum transmembrane protein with thiol-disulfide oxidase activity
Article Snippet: SDS-PAGE and immunoblot detection of PDI8 was performed as described above, with each lane loaded with 20 μL of sample (~7.2 μg of microsomal protein). .. A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO). .. The PDI8 abb’ gene fragment was amplified from a full-length PDI8 cDNA using a forward primer with an XmaI site (5’-TGT CCC GGG AGA TGA TCA ATT CAC CCT CGA C-3’) and a reverse primer with a SalI site (5’-AAT GTC GAC CAT TGA GTT GAT AAA TCC CAT G-3’).

Article Title: Rotavirus Replication Requires a Functional Proteasome for Effective Assembly of Viroplasms
Article Snippet: In order to construct the plasmid pVAX-T7-segment11-ribo-T7stop, indicated in the Results section as pVAX-segment 11, the pVAX vector (Invitrogen) was modified by deletion of the cytomegalovirus (CMV) promoter using NdeI and ApaI restriction enzymes and subsequent insertion of an oligonucleotide containing the XmaI, ApaI, and SacII restriction sites (5′-TATGGGTACCTACTCCCGGGTTACAGGGCCCGCGGAGGCC-3′; Sigma). .. In order to construct the plasmid pVAX-T7-segment11-ribo-T7stop, indicated in the Results section as pVAX-segment 11, the pVAX vector (Invitrogen) was modified by deletion of the cytomegalovirus (CMV) promoter using NdeI and ApaI restriction enzymes and subsequent insertion of an oligonucleotide containing the XmaI, ApaI, and SacII restriction sites (5′-TATGGGTACCTACTCCCGGGTTACAGGGCCCGCGGAGGCC-3′; Sigma).

Cell Culture:

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The protein expression vector for Drosophila cell culture, pAc5.1/TCF/Pan-V5/His (pAc-TCF/Pan), has been described previously . .. The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites.

Generated:

Article Title: Arabidopsis protein disulfide isomerase-8 is a type I endoplasmic reticulum transmembrane protein with thiol-disulfide oxidase activity
Article Snippet: SDS-PAGE and immunoblot detection of PDI8 was performed as described above, with each lane loaded with 20 μL of sample (~7.2 μg of microsomal protein). .. A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO). .. The PDI8 abb’ gene fragment was amplified from a full-length PDI8 cDNA using a forward primer with an XmaI site (5’-TGT CCC GGG AGA TGA TCA ATT CAC CCT CGA C-3’) and a reverse primer with a SalI site (5’-AAT GTC GAC CAT TGA GTT GAT AAA TCC CAT G-3’).

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The Quikchange II mutagenesis kit (Stratagene) was used to generate the various C-clamp mutants in the pAc-TCF/Pan expression vector. pAc-Arm* and the luciferase reporters, pGL3-nkd-intE and pGL3-6xHMG, have been described previously , . .. The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites. .. The pET vector encodes a C terminus 10xHis tag, which was used for protein purification.

Polymerase Chain Reaction:

Article Title: Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine
Article Snippet: Despite this high degree of structural conservation, a marked specificity for the synthesis of A-PG or L-PG was observed for A-PGS and L-PGS in enzymatic assays using Ala-tRNAAla or Lys-tRNALys as substrates, respectively ( ). .. Base pairs 1627–2643 of ORF PA0920 from P . aeruginosa PAO1 and base pairs 1555–2550 of ORF yfiX from B . licheniformis DSM13 were PCR-amplified using oligonucleotide pairs 1 and 2 and 3 and 4 , respectively, and were cloned into the XmaI and SacI sites of pET52b(+) (Novagen) for expression with a cleavable N-terminal Strep -II-tag. .. The SerP (surface entropy reduction) server ( ) was used to identify mutations that may facilitate optimized crystallization of A-PGS (KGKE674 to AGAA674 ).

Recombinant:

Article Title: Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine
Article Snippet: Base pairs 1627–2643 of ORF PA0920 from P . aeruginosa PAO1 and base pairs 1555–2550 of ORF yfiX from B . licheniformis DSM13 were PCR-amplified using oligonucleotide pairs 1 and 2 and 3 and 4 , respectively, and were cloned into the XmaI and SacI sites of pET52b(+) (Novagen) for expression with a cleavable N-terminal Strep -II-tag. .. Base pairs 1627–2643 of ORF PA0920 from P . aeruginosa PAO1 and base pairs 1555–2550 of ORF yfiX from B . licheniformis DSM13 were PCR-amplified using oligonucleotide pairs 1 and 2 and 3 and 4 , respectively, and were cloned into the XmaI and SacI sites of pET52b(+) (Novagen) for expression with a cleavable N-terminal Strep -II-tag.

Mutagenesis:

Article Title: Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine
Article Snippet: Base pairs 1627–2643 of ORF PA0920 from P . aeruginosa PAO1 and base pairs 1555–2550 of ORF yfiX from B . licheniformis DSM13 were PCR-amplified using oligonucleotide pairs 1 and 2 and 3 and 4 , respectively, and were cloned into the XmaI and SacI sites of pET52b(+) (Novagen) for expression with a cleavable N-terminal Strep -II-tag. .. A-PGS and L-PGS genes were expressed, and recombinant proteins were purified to apparent homogeneity as follows: Transformed Escherichia coli BL 21(DE3) cells for the production of A-PGS543–881 and Tuner (DE3) cells for production of L-PGS519–850 were cultivated at 37 °C in LB medium supplemented with 100 µg/mL ampicillin.

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The Quikchange II mutagenesis kit (Stratagene) was used to generate the various C-clamp mutants in the pAc-TCF/Pan expression vector. pAc-Arm* and the luciferase reporters, pGL3-nkd-intE and pGL3-6xHMG, have been described previously , . .. The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites.

Purification:

Article Title: Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine
Article Snippet: Paragraph title: Production and Purification of P . aeruginosa A-PGS and B . licheniformis L-PGS. ... Base pairs 1627–2643 of ORF PA0920 from P . aeruginosa PAO1 and base pairs 1555–2550 of ORF yfiX from B . licheniformis DSM13 were PCR-amplified using oligonucleotide pairs 1 and 2 and 3 and 4 , respectively, and were cloned into the XmaI and SacI sites of pET52b(+) (Novagen) for expression with a cleavable N-terminal Strep -II-tag.

Transgenic Assay:

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites. .. The pET vector encodes a C terminus 10xHis tag, which was used for protein purification.

Plasmid Preparation:

Article Title: Arabidopsis protein disulfide isomerase-8 is a type I endoplasmic reticulum transmembrane protein with thiol-disulfide oxidase activity
Article Snippet: SDS-PAGE and immunoblot detection of PDI8 was performed as described above, with each lane loaded with 20 μL of sample (~7.2 μg of microsomal protein). .. A construct for the heterologous expression of PDI8 in E. coli was generated by cloning the coding sequence for the lumenal portion of PDI8, containing the catalytic a domain and redox-inactive b and b’ domains (PDI8abb’ , Fig. ) between the XmaI and SalI restriction sites of the bacterial expression vector pFLAG-CTS (Sigma-Aldrich, St. Louis, MO). .. The PDI8 abb’ gene fragment was amplified from a full-length PDI8 cDNA using a forward primer with an XmaI site (5’-TGT CCC GGG AGA TGA TCA ATT CAC CCT CGA C-3’) and a reverse primer with a SalI site (5’-AAT GTC GAC CAT TGA GTT GAT AAA TCC CAT G-3’).

Article Title: Structural basis for ubiquitin-mediated antiviral signal activation by RIG-I
Article Snippet: Paragraph title: Plasmid constructs ... Wild type and variants of RIG-I 2CARD (residue 1–200) were subcloned between XmaI and HindIII restriction sites in pET47b (Novagen).

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The Quikchange II mutagenesis kit (Stratagene) was used to generate the various C-clamp mutants in the pAc-TCF/Pan expression vector. pAc-Arm* and the luciferase reporters, pGL3-nkd-intE and pGL3-6xHMG, have been described previously , . .. The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites. .. The pET vector encodes a C terminus 10xHis tag, which was used for protein purification.

Article Title: Rotavirus Replication Requires a Functional Proteasome for Effective Assembly of Viroplasms
Article Snippet: The NSP5 gene was derived from the OSU rotavirus strain, and the NSP2, VP1, VP2 and VP6 genes came from the SA11 strain. .. In order to construct the plasmid pVAX-T7-segment11-ribo-T7stop, indicated in the Results section as pVAX-segment 11, the pVAX vector (Invitrogen) was modified by deletion of the cytomegalovirus (CMV) promoter using NdeI and ApaI restriction enzymes and subsequent insertion of an oligonucleotide containing the XmaI, ApaI, and SacII restriction sites (5′-TATGGGTACCTACTCCCGGGTTACAGGGCCCGCGGAGGCC-3′; Sigma). .. Between the sites of XmaI and ApaI, a synthetic sequence (Genscript) has been inserted in order to clone the ribozyme and the T7 terminator (5′- ccCGGG TCGGCATGGCATCTCCACCTCCTCGCGGTCCGACCTGGGCATCCGAAGGAGGACGTCGTCCACTCGGATGGCTAAGGGAGAGCTcggatccGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATCGAGATCCtctaga gggccc -3′; capital letters indicate the ribozyme, and the T7 terminator and the XmaI and ApaI sites are underlined).

Positron Emission Tomography:

Article Title: Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/ss-catenin Signaling
Article Snippet: The Quikchange II mutagenesis kit (Stratagene) was used to generate the various C-clamp mutants in the pAc-TCF/Pan expression vector. pAc-Arm* and the luciferase reporters, pGL3-nkd-intE and pGL3-6xHMG, have been described previously , . .. The protein expression vectors for EMSA were generated by cloning the region encoding the HMG domain and the C-clamp from the pAc-TCF/Pan constructs into the XmaI and SacI restriction sites of the pET52b(+) vector (pET) (Merck Millipore). pET-HMG and pET-C-clamp were generated by cloning the respective coding regions (residues 271 to 369 for the HMG domain; residues 363 to 408 for the C-clamp) into the same sites. .. The pET vector encodes a C terminus 10xHis tag, which was used for protein purification.

Produced:

Article Title: Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine
Article Snippet: Base pairs 1627–2643 of ORF PA0920 from P . aeruginosa PAO1 and base pairs 1555–2550 of ORF yfiX from B . licheniformis DSM13 were PCR-amplified using oligonucleotide pairs 1 and 2 and 3 and 4 , respectively, and were cloned into the XmaI and SacI sites of pET52b(+) (Novagen) for expression with a cleavable N-terminal Strep -II-tag. .. A-PGS and L-PGS genes were expressed, and recombinant proteins were purified to apparent homogeneity as follows: Transformed Escherichia coli BL 21(DE3) cells for the production of A-PGS543–881 and Tuner (DE3) cells for production of L-PGS519–850 were cultivated at 37 °C in LB medium supplemented with 100 µg/mL ampicillin.

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    Millipore xmai
    Xmai, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    xmai - by Bioz Stars, 2019-12
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