Structured Review

Millipore pet 44a
Pet 44a, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pet 44a/product/Millipore
Average 85 stars, based on 7 article reviews
Price from $9.99 to $1999.99
pet 44a - by Bioz Stars, 2020-08
85/100 stars

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Related Articles

Clone Assay:

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The amplified β2-GP I products were digested with HindIII and BamHI restriction enzymes (Takara Bio, Inc.) and cloned into multiple clone sites of pET-44a (+) (Novagen; Merck KGaA). .. In order to construct the prokaryotic expression vector, the structural gene of human β2-GP I protein was amplified by PCR and inserted into the pET-44a (+) plasmid.

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The amplified β2-GP I products were digested with HindIII and BamHI restriction enzymes (Takara Bio, Inc.) and cloned into multiple clone sites of pET-44a (+) (Novagen; Merck KGaA). .. Purification of the His-tagged protein In order to construct the prokaryotic expression vector, the structural gene of human β2-GP I protein was amplified by PCR and inserted into the pET-44a (+) plasmid.

Positron Emission Tomography:

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The vector of pET-44a(+) encoding the His-tag sequence and the expression of fusion protein rhβ2-GP I was induced by 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG; Sigma-Aldrich; Merck KGaA); the bacteria were incubated at 25°C, 100 rpm/min for 2, 4 or 6 h. Subsequently, the bacteria was lysed with lysis buffer (50 mM NaH2 PO4 , 300 mM NaCl, 5 mM imidazole and 1 mg/ml lysozyme, pH=8.0) then sonicated 6 times at 200 W in 10 sec bursts with a 10 sec cooling period between each burst. .. The induced protein in the lysate was further purified by Ni-NTA agarose (Qiagen, Germantown, MD, USA) and identified by western blotting as described below.

Article Title: Structure-based analysis of the guanine nucleotide exchange factor SmgGDS reveals armadillo-repeat motifs and key regions for activity and GTPase binding
Article Snippet: .. The following cDNA were subcloned into the pET-44a(+) vector (Novagen): h RhoA (UniProt ID: P61586-1, aa 1–193) harbored His6 and PreScission protease recognition sequences at the N terminus. h RhoAL193A was generated by PCR-based site-directed mutagenesis. .. The host cell and protein expression procedure were the same as described above.

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The amplified β2-GP I products were digested with HindIII and BamHI restriction enzymes (Takara Bio, Inc.) and cloned into multiple clone sites of pET-44a (+) (Novagen; Merck KGaA). .. In order to construct the prokaryotic expression vector, the structural gene of human β2-GP I protein was amplified by PCR and inserted into the pET-44a (+) plasmid.

Article Title: Homologous high-throughput expression and purification of highly conserved E coli proteins
Article Snippet: .. Destination vector pD-Nus1 [GenBank: EF431918 ] was derived from pET-44a (Novagen). .. Vector pET-44a was digested with SacII and XhoI.

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The amplified β2-GP I products were digested with HindIII and BamHI restriction enzymes (Takara Bio, Inc.) and cloned into multiple clone sites of pET-44a (+) (Novagen; Merck KGaA). .. Purification of the His-tagged protein In order to construct the prokaryotic expression vector, the structural gene of human β2-GP I protein was amplified by PCR and inserted into the pET-44a (+) plasmid.

Article Title: Functional Importance of the Anaphase-Promoting Complex-Cdh1-Mediated Degradation of TMAP/CKAP2 in Regulation of Spindle Function and Cytokinesis ▿Functional Importance of the Anaphase-Promoting Complex-Cdh1-Mediated Degradation of TMAP/CKAP2 in Regulation of Spindle Function and Cytokinesis ▿ †
Article Snippet: .. For production of Nus-tagged TMAP/CKAP2 (Nus-CKAP2), the full-length coding sequence of TMAP/CKAP2 was subcloned into pET-44a(+) vector (Novagen). .. It was introduced to and expressed in an Escherichia coli strain BL21 (Novagen) according to the manufacturer's instructions.

Mutagenesis:

Article Title: Structure-based analysis of the guanine nucleotide exchange factor SmgGDS reveals armadillo-repeat motifs and key regions for activity and GTPase binding
Article Snippet: .. The following cDNA were subcloned into the pET-44a(+) vector (Novagen): h RhoA (UniProt ID: P61586-1, aa 1–193) harbored His6 and PreScission protease recognition sequences at the N terminus. h RhoAL193A was generated by PCR-based site-directed mutagenesis. .. The host cell and protein expression procedure were the same as described above.

Size-exclusion Chromatography:

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The vector of pET-44a(+) encoding the His-tag sequence and the expression of fusion protein rhβ2-GP I was induced by 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG; Sigma-Aldrich; Merck KGaA); the bacteria were incubated at 25°C, 100 rpm/min for 2, 4 or 6 h. Subsequently, the bacteria was lysed with lysis buffer (50 mM NaH2 PO4 , 300 mM NaCl, 5 mM imidazole and 1 mg/ml lysozyme, pH=8.0) then sonicated 6 times at 200 W in 10 sec bursts with a 10 sec cooling period between each burst. .. The induced protein in the lysate was further purified by Ni-NTA agarose (Qiagen, Germantown, MD, USA) and identified by western blotting as described below.

Sequencing:

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The vector of pET-44a(+) encoding the His-tag sequence and the expression of fusion protein rhβ2-GP I was induced by 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG; Sigma-Aldrich; Merck KGaA); the bacteria were incubated at 25°C, 100 rpm/min for 2, 4 or 6 h. Subsequently, the bacteria was lysed with lysis buffer (50 mM NaH2 PO4 , 300 mM NaCl, 5 mM imidazole and 1 mg/ml lysozyme, pH=8.0) then sonicated 6 times at 200 W in 10 sec bursts with a 10 sec cooling period between each burst. .. The induced protein in the lysate was further purified by Ni-NTA agarose (Qiagen, Germantown, MD, USA) and identified by western blotting as described below.

Article Title: Functional Importance of the Anaphase-Promoting Complex-Cdh1-Mediated Degradation of TMAP/CKAP2 in Regulation of Spindle Function and Cytokinesis ▿Functional Importance of the Anaphase-Promoting Complex-Cdh1-Mediated Degradation of TMAP/CKAP2 in Regulation of Spindle Function and Cytokinesis ▿ †
Article Snippet: .. For production of Nus-tagged TMAP/CKAP2 (Nus-CKAP2), the full-length coding sequence of TMAP/CKAP2 was subcloned into pET-44a(+) vector (Novagen). .. It was introduced to and expressed in an Escherichia coli strain BL21 (Novagen) according to the manufacturer's instructions.

Incubation:

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The vector of pET-44a(+) encoding the His-tag sequence and the expression of fusion protein rhβ2-GP I was induced by 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG; Sigma-Aldrich; Merck KGaA); the bacteria were incubated at 25°C, 100 rpm/min for 2, 4 or 6 h. Subsequently, the bacteria was lysed with lysis buffer (50 mM NaH2 PO4 , 300 mM NaCl, 5 mM imidazole and 1 mg/ml lysozyme, pH=8.0) then sonicated 6 times at 200 W in 10 sec bursts with a 10 sec cooling period between each burst. .. The induced protein in the lysate was further purified by Ni-NTA agarose (Qiagen, Germantown, MD, USA) and identified by western blotting as described below.

Amplification:

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The amplified β2-GP I products were digested with HindIII and BamHI restriction enzymes (Takara Bio, Inc.) and cloned into multiple clone sites of pET-44a (+) (Novagen; Merck KGaA). .. In order to construct the prokaryotic expression vector, the structural gene of human β2-GP I protein was amplified by PCR and inserted into the pET-44a (+) plasmid.

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The amplified β2-GP I products were digested with HindIII and BamHI restriction enzymes (Takara Bio, Inc.) and cloned into multiple clone sites of pET-44a (+) (Novagen; Merck KGaA). .. Purification of the His-tagged protein In order to construct the prokaryotic expression vector, the structural gene of human β2-GP I protein was amplified by PCR and inserted into the pET-44a (+) plasmid.

Plasmid Preparation:

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The vector of pET-44a(+) encoding the His-tag sequence and the expression of fusion protein rhβ2-GP I was induced by 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG; Sigma-Aldrich; Merck KGaA); the bacteria were incubated at 25°C, 100 rpm/min for 2, 4 or 6 h. Subsequently, the bacteria was lysed with lysis buffer (50 mM NaH2 PO4 , 300 mM NaCl, 5 mM imidazole and 1 mg/ml lysozyme, pH=8.0) then sonicated 6 times at 200 W in 10 sec bursts with a 10 sec cooling period between each burst. .. The induced protein in the lysate was further purified by Ni-NTA agarose (Qiagen, Germantown, MD, USA) and identified by western blotting as described below.

Article Title: Structure-based analysis of the guanine nucleotide exchange factor SmgGDS reveals armadillo-repeat motifs and key regions for activity and GTPase binding
Article Snippet: .. The following cDNA were subcloned into the pET-44a(+) vector (Novagen): h RhoA (UniProt ID: P61586-1, aa 1–193) harbored His6 and PreScission protease recognition sequences at the N terminus. h RhoAL193A was generated by PCR-based site-directed mutagenesis. .. The host cell and protein expression procedure were the same as described above.

Article Title: Homologous high-throughput expression and purification of highly conserved E coli proteins
Article Snippet: .. Destination vector pD-Nus1 [GenBank: EF431918 ] was derived from pET-44a (Novagen). .. Vector pET-44a was digested with SacII and XhoI.

Article Title: Functional Importance of the Anaphase-Promoting Complex-Cdh1-Mediated Degradation of TMAP/CKAP2 in Regulation of Spindle Function and Cytokinesis ▿Functional Importance of the Anaphase-Promoting Complex-Cdh1-Mediated Degradation of TMAP/CKAP2 in Regulation of Spindle Function and Cytokinesis ▿ †
Article Snippet: .. For production of Nus-tagged TMAP/CKAP2 (Nus-CKAP2), the full-length coding sequence of TMAP/CKAP2 was subcloned into pET-44a(+) vector (Novagen). .. It was introduced to and expressed in an Escherichia coli strain BL21 (Novagen) according to the manufacturer's instructions.

Expressing:

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The vector of pET-44a(+) encoding the His-tag sequence and the expression of fusion protein rhβ2-GP I was induced by 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG; Sigma-Aldrich; Merck KGaA); the bacteria were incubated at 25°C, 100 rpm/min for 2, 4 or 6 h. Subsequently, the bacteria was lysed with lysis buffer (50 mM NaH2 PO4 , 300 mM NaCl, 5 mM imidazole and 1 mg/ml lysozyme, pH=8.0) then sonicated 6 times at 200 W in 10 sec bursts with a 10 sec cooling period between each burst. .. The induced protein in the lysate was further purified by Ni-NTA agarose (Qiagen, Germantown, MD, USA) and identified by western blotting as described below.

Lysis:

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The vector of pET-44a(+) encoding the His-tag sequence and the expression of fusion protein rhβ2-GP I was induced by 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG; Sigma-Aldrich; Merck KGaA); the bacteria were incubated at 25°C, 100 rpm/min for 2, 4 or 6 h. Subsequently, the bacteria was lysed with lysis buffer (50 mM NaH2 PO4 , 300 mM NaCl, 5 mM imidazole and 1 mg/ml lysozyme, pH=8.0) then sonicated 6 times at 200 W in 10 sec bursts with a 10 sec cooling period between each burst. .. The induced protein in the lysate was further purified by Ni-NTA agarose (Qiagen, Germantown, MD, USA) and identified by western blotting as described below.

Polymerase Chain Reaction:

Article Title: Structure-based analysis of the guanine nucleotide exchange factor SmgGDS reveals armadillo-repeat motifs and key regions for activity and GTPase binding
Article Snippet: .. The following cDNA were subcloned into the pET-44a(+) vector (Novagen): h RhoA (UniProt ID: P61586-1, aa 1–193) harbored His6 and PreScission protease recognition sequences at the N terminus. h RhoAL193A was generated by PCR-based site-directed mutagenesis. .. The host cell and protein expression procedure were the same as described above.

Sonication:

Article Title: Effect and mechanism of the aβ2-GP I/rhβ2-GP I complex on JEG-3 cell proliferation, migration and invasion
Article Snippet: .. The vector of pET-44a(+) encoding the His-tag sequence and the expression of fusion protein rhβ2-GP I was induced by 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG; Sigma-Aldrich; Merck KGaA); the bacteria were incubated at 25°C, 100 rpm/min for 2, 4 or 6 h. Subsequently, the bacteria was lysed with lysis buffer (50 mM NaH2 PO4 , 300 mM NaCl, 5 mM imidazole and 1 mg/ml lysozyme, pH=8.0) then sonicated 6 times at 200 W in 10 sec bursts with a 10 sec cooling period between each burst. .. The induced protein in the lysate was further purified by Ni-NTA agarose (Qiagen, Germantown, MD, USA) and identified by western blotting as described below.

Derivative Assay:

Article Title: Homologous high-throughput expression and purification of highly conserved E coli proteins
Article Snippet: .. Destination vector pD-Nus1 [GenBank: EF431918 ] was derived from pET-44a (Novagen). .. Vector pET-44a was digested with SacII and XhoI.

Generated:

Article Title: Structure-based analysis of the guanine nucleotide exchange factor SmgGDS reveals armadillo-repeat motifs and key regions for activity and GTPase binding
Article Snippet: .. The following cDNA were subcloned into the pET-44a(+) vector (Novagen): h RhoA (UniProt ID: P61586-1, aa 1–193) harbored His6 and PreScission protease recognition sequences at the N terminus. h RhoAL193A was generated by PCR-based site-directed mutagenesis. .. The host cell and protein expression procedure were the same as described above.

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    Millipore 42a 44a
    42a 44a, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/42a 44a/product/Millipore
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    42a 44a - by Bioz Stars, 2020-08
    86/100 stars
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