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  • 85
    GE Healthcare ge healthcare pgex
    Ge Healthcare Pgex, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 6p1 ge healthcare
    Pgex 6p1 Ge Healthcare, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 84/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 6 1 vector ge healthcare
    Pgex 6 1 Vector Ge Healthcare, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 4t1
    Expression of the Be82 gene products in E. coli cells stained with Coomassie blue. (A) Lysates of cells with the pGEMEX-2 (lane a) and pGEMEX-2/Be82 (lane b) expression vectors; (B) lysates of cells with the <t>pGEX-4T</t> (lane a) and pGEX-4T/Be82 (lane b) expression vectors. Molecular size markers (in kilodaltons) are shown to the left and right of the lanes.
    Pgex 4t1, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 94/100, based on 1719 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 2t
    Western-blot analysis of the SCP2 domain Left panel: proteins from various tissues within the last larval instar were used. Lane 1, midgut; lane 2, fat body; lane 3, Malpighian tubules; lane 4, recombinant protein corresponding to the SCP2 domain of SCPx. The antibody against the SCP2 domain of S. littoralis SCPx protein was used. Right panel: Western-blot analysis of the recombinant S. littoralis SCPx protein using anti-(SCP2 domain) antibody. Lane 1, proteins from E. coli carrying the <t>pGEX-2T</t> vector without the SCPx cDNA; lane 2, proteins from E. coli carrying the pGEX-2T vector containing SCPx cDNA with induction of expression.
    Pgex 2t, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 2186 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 2tk
    Schematic diagram of experimental approach. A PCR product containing 17 random codons was inserted into the BamHI site of <t>pGEX-2TK</t> producing various glutathione-S-transferase fusion proteins which were bound to glutathione-sepharose, and labeled with 32 P using protein kinase A. After washing and thrombin digestion, the labeled peptides were incubated with several different cell lines and assayed.
    Pgex 2tk, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 90/100, based on 663 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 6p2
    Schematic diagram of experimental approach. A PCR product containing 17 random codons was inserted into the BamHI site of <t>pGEX-2TK</t> producing various glutathione-S-transferase fusion proteins which were bound to glutathione-sepharose, and labeled with 32 P using protein kinase A. After washing and thrombin digestion, the labeled peptides were incubated with several different cell lines and assayed.
    Pgex 6p2, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 91/100, based on 225 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 6p 3
    Establishment of monkey podoplanin-neutralizing antibodies (A) Purification of a monkey podoplanin (mkyPDPN) immunogen to establish a hybridoma secreting anti-PDPN monoclonal antibodies (mAbs). A mkyPDPN cDNA region encoding amino acids 76–89 (226–267 bp) was tandemly connected 21 times. The cDNA fragment was inserted into a <t>pGEX-6P-3</t> vector, and a glutathione S-transferase (GST)-tagged mkyPDPN peptide (76–89 aa) produced by BL21 (DE3) E. coli was purified using glutathione sepharose. BALB/c mice were injected with the GST-tagged peptide, after which their splenocytes were fused with mouse myeloma P3U1 cells using polyethylene glycol. Hybridoma screening and antibody purification from ascites were performed. (B) CHO cells transfected with an empty vector (mock), wild-type monkey podoplanin (mkyPDPN-WT), wild-type human podoplanin (hPDPN-WT) were treated with PBS (closed areas) or antibodies (open areas), including anti-PDPN mAb D2-40, 1F6, 2F7, or 3F4 to measure PDPN expression levels. (C) GST-tagged recombinant mkyPDPN protein (WT) and its point mutants were expressed in E. coli . Cell lysates were electrophoresed and immunoblotted with antibodies to PDPN (1F6, 2F7, 3F4) or GST. The PLAG4 domain is indicated by red letters. (D) CHO/mock, CHO/mkyPDPN-WT, or CHO/hPDPN-WT cells were incubated with 100 μg/mL of control IgG1 or anti-PDPN antibodies 1F6, 2F7, or 3F4, followed by incubation with 1 μg/mL of hCLEC-2-(His) 10 (open areas: control IgG-treated samples; green areas: anti-PDPN mAb-treated samples). After washing, cells were further incubated with Alexa Flour 488-conjugated anti-penta-His second antibody. CLEC-2 binding was measured by flow cytometry. Gray areas indicate the fluorescence intensity of samples not treated with CLEC-2. (E) CHO/mkyPDPN-WT cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs followed by incubation with mouse platelet-rich plasma (PRP). The aggregation rate was estimated using an aggregometer. (F) PLAG3 domain-deleted mkyPDPN mutant cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs, followed by incubation with mouse PRP. The aggregation rate was estimated using an aggregometer.
    Pgex 6p 3, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 624 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 5x2
    Establishment of monkey podoplanin-neutralizing antibodies (A) Purification of a monkey podoplanin (mkyPDPN) immunogen to establish a hybridoma secreting anti-PDPN monoclonal antibodies (mAbs). A mkyPDPN cDNA region encoding amino acids 76–89 (226–267 bp) was tandemly connected 21 times. The cDNA fragment was inserted into a <t>pGEX-6P-3</t> vector, and a glutathione S-transferase (GST)-tagged mkyPDPN peptide (76–89 aa) produced by BL21 (DE3) E. coli was purified using glutathione sepharose. BALB/c mice were injected with the GST-tagged peptide, after which their splenocytes were fused with mouse myeloma P3U1 cells using polyethylene glycol. Hybridoma screening and antibody purification from ascites were performed. (B) CHO cells transfected with an empty vector (mock), wild-type monkey podoplanin (mkyPDPN-WT), wild-type human podoplanin (hPDPN-WT) were treated with PBS (closed areas) or antibodies (open areas), including anti-PDPN mAb D2-40, 1F6, 2F7, or 3F4 to measure PDPN expression levels. (C) GST-tagged recombinant mkyPDPN protein (WT) and its point mutants were expressed in E. coli . Cell lysates were electrophoresed and immunoblotted with antibodies to PDPN (1F6, 2F7, 3F4) or GST. The PLAG4 domain is indicated by red letters. (D) CHO/mock, CHO/mkyPDPN-WT, or CHO/hPDPN-WT cells were incubated with 100 μg/mL of control IgG1 or anti-PDPN antibodies 1F6, 2F7, or 3F4, followed by incubation with 1 μg/mL of hCLEC-2-(His) 10 (open areas: control IgG-treated samples; green areas: anti-PDPN mAb-treated samples). After washing, cells were further incubated with Alexa Flour 488-conjugated anti-penta-His second antibody. CLEC-2 binding was measured by flow cytometry. Gray areas indicate the fluorescence intensity of samples not treated with CLEC-2. (E) CHO/mkyPDPN-WT cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs followed by incubation with mouse platelet-rich plasma (PRP). The aggregation rate was estimated using an aggregometer. (F) PLAG3 domain-deleted mkyPDPN mutant cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs, followed by incubation with mouse PRP. The aggregation rate was estimated using an aggregometer.
    Pgex 5x2, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 6p
    Establishment of monkey podoplanin-neutralizing antibodies (A) Purification of a monkey podoplanin (mkyPDPN) immunogen to establish a hybridoma secreting anti-PDPN monoclonal antibodies (mAbs). A mkyPDPN cDNA region encoding amino acids 76–89 (226–267 bp) was tandemly connected 21 times. The cDNA fragment was inserted into a <t>pGEX-6P-3</t> vector, and a glutathione S-transferase (GST)-tagged mkyPDPN peptide (76–89 aa) produced by BL21 (DE3) E. coli was purified using glutathione sepharose. BALB/c mice were injected with the GST-tagged peptide, after which their splenocytes were fused with mouse myeloma P3U1 cells using polyethylene glycol. Hybridoma screening and antibody purification from ascites were performed. (B) CHO cells transfected with an empty vector (mock), wild-type monkey podoplanin (mkyPDPN-WT), wild-type human podoplanin (hPDPN-WT) were treated with PBS (closed areas) or antibodies (open areas), including anti-PDPN mAb D2-40, 1F6, 2F7, or 3F4 to measure PDPN expression levels. (C) GST-tagged recombinant mkyPDPN protein (WT) and its point mutants were expressed in E. coli . Cell lysates were electrophoresed and immunoblotted with antibodies to PDPN (1F6, 2F7, 3F4) or GST. The PLAG4 domain is indicated by red letters. (D) CHO/mock, CHO/mkyPDPN-WT, or CHO/hPDPN-WT cells were incubated with 100 μg/mL of control IgG1 or anti-PDPN antibodies 1F6, 2F7, or 3F4, followed by incubation with 1 μg/mL of hCLEC-2-(His) 10 (open areas: control IgG-treated samples; green areas: anti-PDPN mAb-treated samples). After washing, cells were further incubated with Alexa Flour 488-conjugated anti-penta-His second antibody. CLEC-2 binding was measured by flow cytometry. Gray areas indicate the fluorescence intensity of samples not treated with CLEC-2. (E) CHO/mkyPDPN-WT cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs followed by incubation with mouse platelet-rich plasma (PRP). The aggregation rate was estimated using an aggregometer. (F) PLAG3 domain-deleted mkyPDPN mutant cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs, followed by incubation with mouse PRP. The aggregation rate was estimated using an aggregometer.
    Pgex 6p, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 89/100, based on 500 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex gst
    Establishment of monkey podoplanin-neutralizing antibodies (A) Purification of a monkey podoplanin (mkyPDPN) immunogen to establish a hybridoma secreting anti-PDPN monoclonal antibodies (mAbs). A mkyPDPN cDNA region encoding amino acids 76–89 (226–267 bp) was tandemly connected 21 times. The cDNA fragment was inserted into a <t>pGEX-6P-3</t> vector, and a glutathione S-transferase (GST)-tagged mkyPDPN peptide (76–89 aa) produced by BL21 (DE3) E. coli was purified using glutathione sepharose. BALB/c mice were injected with the GST-tagged peptide, after which their splenocytes were fused with mouse myeloma P3U1 cells using polyethylene glycol. Hybridoma screening and antibody purification from ascites were performed. (B) CHO cells transfected with an empty vector (mock), wild-type monkey podoplanin (mkyPDPN-WT), wild-type human podoplanin (hPDPN-WT) were treated with PBS (closed areas) or antibodies (open areas), including anti-PDPN mAb D2-40, 1F6, 2F7, or 3F4 to measure PDPN expression levels. (C) GST-tagged recombinant mkyPDPN protein (WT) and its point mutants were expressed in E. coli . Cell lysates were electrophoresed and immunoblotted with antibodies to PDPN (1F6, 2F7, 3F4) or GST. The PLAG4 domain is indicated by red letters. (D) CHO/mock, CHO/mkyPDPN-WT, or CHO/hPDPN-WT cells were incubated with 100 μg/mL of control IgG1 or anti-PDPN antibodies 1F6, 2F7, or 3F4, followed by incubation with 1 μg/mL of hCLEC-2-(His) 10 (open areas: control IgG-treated samples; green areas: anti-PDPN mAb-treated samples). After washing, cells were further incubated with Alexa Flour 488-conjugated anti-penta-His second antibody. CLEC-2 binding was measured by flow cytometry. Gray areas indicate the fluorescence intensity of samples not treated with CLEC-2. (E) CHO/mkyPDPN-WT cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs followed by incubation with mouse platelet-rich plasma (PRP). The aggregation rate was estimated using an aggregometer. (F) PLAG3 domain-deleted mkyPDPN mutant cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs, followed by incubation with mouse PRP. The aggregation rate was estimated using an aggregometer.
    Pgex Gst, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex kg
    Establishment of monkey podoplanin-neutralizing antibodies (A) Purification of a monkey podoplanin (mkyPDPN) immunogen to establish a hybridoma secreting anti-PDPN monoclonal antibodies (mAbs). A mkyPDPN cDNA region encoding amino acids 76–89 (226–267 bp) was tandemly connected 21 times. The cDNA fragment was inserted into a <t>pGEX-6P-3</t> vector, and a glutathione S-transferase (GST)-tagged mkyPDPN peptide (76–89 aa) produced by BL21 (DE3) E. coli was purified using glutathione sepharose. BALB/c mice were injected with the GST-tagged peptide, after which their splenocytes were fused with mouse myeloma P3U1 cells using polyethylene glycol. Hybridoma screening and antibody purification from ascites were performed. (B) CHO cells transfected with an empty vector (mock), wild-type monkey podoplanin (mkyPDPN-WT), wild-type human podoplanin (hPDPN-WT) were treated with PBS (closed areas) or antibodies (open areas), including anti-PDPN mAb D2-40, 1F6, 2F7, or 3F4 to measure PDPN expression levels. (C) GST-tagged recombinant mkyPDPN protein (WT) and its point mutants were expressed in E. coli . Cell lysates were electrophoresed and immunoblotted with antibodies to PDPN (1F6, 2F7, 3F4) or GST. The PLAG4 domain is indicated by red letters. (D) CHO/mock, CHO/mkyPDPN-WT, or CHO/hPDPN-WT cells were incubated with 100 μg/mL of control IgG1 or anti-PDPN antibodies 1F6, 2F7, or 3F4, followed by incubation with 1 μg/mL of hCLEC-2-(His) 10 (open areas: control IgG-treated samples; green areas: anti-PDPN mAb-treated samples). After washing, cells were further incubated with Alexa Flour 488-conjugated anti-penta-His second antibody. CLEC-2 binding was measured by flow cytometry. Gray areas indicate the fluorescence intensity of samples not treated with CLEC-2. (E) CHO/mkyPDPN-WT cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs followed by incubation with mouse platelet-rich plasma (PRP). The aggregation rate was estimated using an aggregometer. (F) PLAG3 domain-deleted mkyPDPN mutant cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs, followed by incubation with mouse PRP. The aggregation rate was estimated using an aggregometer.
    Pgex Kg, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 91/100, based on 555 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 5x
    Establishment of monkey podoplanin-neutralizing antibodies (A) Purification of a monkey podoplanin (mkyPDPN) immunogen to establish a hybridoma secreting anti-PDPN monoclonal antibodies (mAbs). A mkyPDPN cDNA region encoding amino acids 76–89 (226–267 bp) was tandemly connected 21 times. The cDNA fragment was inserted into a <t>pGEX-6P-3</t> vector, and a glutathione S-transferase (GST)-tagged mkyPDPN peptide (76–89 aa) produced by BL21 (DE3) E. coli was purified using glutathione sepharose. BALB/c mice were injected with the GST-tagged peptide, after which their splenocytes were fused with mouse myeloma P3U1 cells using polyethylene glycol. Hybridoma screening and antibody purification from ascites were performed. (B) CHO cells transfected with an empty vector (mock), wild-type monkey podoplanin (mkyPDPN-WT), wild-type human podoplanin (hPDPN-WT) were treated with PBS (closed areas) or antibodies (open areas), including anti-PDPN mAb D2-40, 1F6, 2F7, or 3F4 to measure PDPN expression levels. (C) GST-tagged recombinant mkyPDPN protein (WT) and its point mutants were expressed in E. coli . Cell lysates were electrophoresed and immunoblotted with antibodies to PDPN (1F6, 2F7, 3F4) or GST. The PLAG4 domain is indicated by red letters. (D) CHO/mock, CHO/mkyPDPN-WT, or CHO/hPDPN-WT cells were incubated with 100 μg/mL of control IgG1 or anti-PDPN antibodies 1F6, 2F7, or 3F4, followed by incubation with 1 μg/mL of hCLEC-2-(His) 10 (open areas: control IgG-treated samples; green areas: anti-PDPN mAb-treated samples). After washing, cells were further incubated with Alexa Flour 488-conjugated anti-penta-His second antibody. CLEC-2 binding was measured by flow cytometry. Gray areas indicate the fluorescence intensity of samples not treated with CLEC-2. (E) CHO/mkyPDPN-WT cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs followed by incubation with mouse platelet-rich plasma (PRP). The aggregation rate was estimated using an aggregometer. (F) PLAG3 domain-deleted mkyPDPN mutant cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs, followed by incubation with mouse PRP. The aggregation rate was estimated using an aggregometer.
    Pgex 5x, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 190 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 3
    Establishment of monkey podoplanin-neutralizing antibodies (A) Purification of a monkey podoplanin (mkyPDPN) immunogen to establish a hybridoma secreting anti-PDPN monoclonal antibodies (mAbs). A mkyPDPN cDNA region encoding amino acids 76–89 (226–267 bp) was tandemly connected 21 times. The cDNA fragment was inserted into a <t>pGEX-6P-3</t> vector, and a glutathione S-transferase (GST)-tagged mkyPDPN peptide (76–89 aa) produced by BL21 (DE3) E. coli was purified using glutathione sepharose. BALB/c mice were injected with the GST-tagged peptide, after which their splenocytes were fused with mouse myeloma P3U1 cells using polyethylene glycol. Hybridoma screening and antibody purification from ascites were performed. (B) CHO cells transfected with an empty vector (mock), wild-type monkey podoplanin (mkyPDPN-WT), wild-type human podoplanin (hPDPN-WT) were treated with PBS (closed areas) or antibodies (open areas), including anti-PDPN mAb D2-40, 1F6, 2F7, or 3F4 to measure PDPN expression levels. (C) GST-tagged recombinant mkyPDPN protein (WT) and its point mutants were expressed in E. coli . Cell lysates were electrophoresed and immunoblotted with antibodies to PDPN (1F6, 2F7, 3F4) or GST. The PLAG4 domain is indicated by red letters. (D) CHO/mock, CHO/mkyPDPN-WT, or CHO/hPDPN-WT cells were incubated with 100 μg/mL of control IgG1 or anti-PDPN antibodies 1F6, 2F7, or 3F4, followed by incubation with 1 μg/mL of hCLEC-2-(His) 10 (open areas: control IgG-treated samples; green areas: anti-PDPN mAb-treated samples). After washing, cells were further incubated with Alexa Flour 488-conjugated anti-penta-His second antibody. CLEC-2 binding was measured by flow cytometry. Gray areas indicate the fluorescence intensity of samples not treated with CLEC-2. (E) CHO/mkyPDPN-WT cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs followed by incubation with mouse platelet-rich plasma (PRP). The aggregation rate was estimated using an aggregometer. (F) PLAG3 domain-deleted mkyPDPN mutant cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs, followed by incubation with mouse PRP. The aggregation rate was estimated using an aggregometer.
    Pgex 3, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 88 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 3x
    Western blot analysis of Itih5 protein with the α-Itih5 antibody. (A) Characterization of the α-Itih5 antibody. Western blot analysis of protein extracts from E. coli strain BL21 producing the <t>pGEX-3X/Itih5</t> recombinant protein. Lane 1: Protein extract without IPTG induction; lane 2: Protein extract with IPTG induction. The arrow indicates the molecular size of the specific signal for the GST-Itih5 protein (approximately 105 kDa). (B) Western blot characterization of the Itih5 isoforms produced in the female genital tract and the liver at E18.5. Arrows indicate the molecular sizes of the specific bands found in the embryonic female genital tract (105 and 50 kDa) and in the liver (73 kDa). (C) Size characterization of the Itih5 isoform produced in the uterus of pregnant and nonpregnant adult mice. The arrow indicates the molecular size of the specific band found in the uterus during gestation and in nonpregnant animals (73 kDa). E, embryonic day; NP, nonpregnant.
    Pgex 3x, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 93/100, based on 740 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 4t3
    Concept of the asymmetric directional T-vector. ( A ) Construction of <t>pGEX-4T3-PRESAT</t> and direct cloning of PCR product in pGEX-4T3-PRESAT. ( B ) The schematic representation of the ORF selection method using potential restriction enzyme site. The figure illustrates the case in which NcoI is chosen as the second restriction enzyme for selection. The rear PCR primer is designed with 5′-GG at the 5′ end, so that only the ligated plasmid with insert in the reverse orientation will have the NcoI site at the TA-cloning position. For NdeI selection, the rear primer with 5′-ATG is used instead of the 5′-GG primer.
    Pgex 4t3, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 93/100, based on 556 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 5x1
    Concept of the asymmetric directional T-vector. ( A ) Construction of <t>pGEX-4T3-PRESAT</t> and direct cloning of PCR product in pGEX-4T3-PRESAT. ( B ) The schematic representation of the ORF selection method using potential restriction enzyme site. The figure illustrates the case in which NcoI is chosen as the second restriction enzyme for selection. The rear PCR primer is designed with 5′-GG at the 5′ end, so that only the ligated plasmid with insert in the reverse orientation will have the NcoI site at the TA-cloning position. For NdeI selection, the rear primer with 5′-ATG is used instead of the 5′-GG primer.
    Pgex 5x1, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 93/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pgex 5
    Concept of the asymmetric directional T-vector. ( A ) Construction of <t>pGEX-4T3-PRESAT</t> and direct cloning of PCR product in pGEX-4T3-PRESAT. ( B ) The schematic representation of the ORF selection method using potential restriction enzyme site. The figure illustrates the case in which NcoI is chosen as the second restriction enzyme for selection. The rear PCR primer is designed with 5′-GG at the 5′ end, so that only the ligated plasmid with insert in the reverse orientation will have the NcoI site at the TA-cloning position. For NdeI selection, the rear primer with 5′-ATG is used instead of the 5′-GG primer.
    Pgex 5, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    GE Healthcare pgex 4t 2
    HCV genome and recombinant proteins. (A) The HCV genome contains a single major open reading frame (ORF) flanked by untranslated regions (UTRs). The 10 proteins encoded within the main ORF are indicated by alternated shading. (B) Genomic fragments of HCV were cloned into the prokaryotic expression vectors, pET32a(+), pQE30, or <t>pGEX-4T-2,</t> in-frame downstream of the 6-His-tag or glutathione S-transferase (GST)-tag coding sequence.
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    Expression of the Be82 gene products in E. coli cells stained with Coomassie blue. (A) Lysates of cells with the pGEMEX-2 (lane a) and pGEMEX-2/Be82 (lane b) expression vectors; (B) lysates of cells with the pGEX-4T (lane a) and pGEX-4T/Be82 (lane b) expression vectors. Molecular size markers (in kilodaltons) are shown to the left and right of the lanes.

    Journal: Journal of Clinical Microbiology

    Article Title: Cloning of a Truncated Babesia equi Gene Encoding an 82-Kilodalton Protein and Its Potential Use in an Enzyme-Linked Immunosorbent Assay

    doi: 10.1128/JCM.40.4.1470-1474.2002

    Figure Lengend Snippet: Expression of the Be82 gene products in E. coli cells stained with Coomassie blue. (A) Lysates of cells with the pGEMEX-2 (lane a) and pGEMEX-2/Be82 (lane b) expression vectors; (B) lysates of cells with the pGEX-4T (lane a) and pGEX-4T/Be82 (lane b) expression vectors. Molecular size markers (in kilodaltons) are shown to the left and right of the lanes.

    Article Snippet: After digestion of pBS/Be82 with Eco RI and Xho I, a 1,953-bp Eco RI and Xho I fragment containing the complete cDNA insert was subcloned into the Eco RI and Xho I sites of the pGEMEX-2 (Promega Corp., Madison, Wis.) and pGEX-4T (Amersham Pharmacia Biotech, Little Chalfont, Buckinghamshire, England) E. coli expression plasmid vectors, and the resulting plasmids were designated pGEMEX-2/Be82 and pGEX/Be82, respectively, and later used to transform the E. coli strain BL21 (Stratagene) according to standard techniques ( ).

    Techniques: Expressing, Staining

    Cloning strategies for cloning the paracoccin ORF for expression. The left panel shows the standard strategy, with PCR amplification of the largest exon, restriction endonuclease digestion, and cloning into the expression vector pGEX-4T-1. The positions of the exons are displayed on the map of the gene. Genomic DNA template was extracted from P. brasiliensis strain Pb18. Agarose gel electrophoresis (mid-left) shows the corresponding band amplified by PCR. The exon 4 amplicon was cloned by Bam HI and Eco RI digestion. The right panel shows the strategy for synthesis of the predicted paracoccin sequence fused with the 5′-UTR elements for transcription in the vector pUC57. Green arrow, T7 promoter; black box, lacO (lac operator); and red box, the phage T7 trailer sequence for ribosome binding.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Recombinant Paracoccin Reproduces the Biological Properties of the Native Protein and Induces Protective Th1 Immunity against Paracoccidioides brasiliensis Infection

    doi: 10.1371/journal.pntd.0002788

    Figure Lengend Snippet: Cloning strategies for cloning the paracoccin ORF for expression. The left panel shows the standard strategy, with PCR amplification of the largest exon, restriction endonuclease digestion, and cloning into the expression vector pGEX-4T-1. The positions of the exons are displayed on the map of the gene. Genomic DNA template was extracted from P. brasiliensis strain Pb18. Agarose gel electrophoresis (mid-left) shows the corresponding band amplified by PCR. The exon 4 amplicon was cloned by Bam HI and Eco RI digestion. The right panel shows the strategy for synthesis of the predicted paracoccin sequence fused with the 5′-UTR elements for transcription in the vector pUC57. Green arrow, T7 promoter; black box, lacO (lac operator); and red box, the phage T7 trailer sequence for ribosome binding.

    Article Snippet: First, the largest exon was amplified from P. brasiliensis genomic DNA using the oligonucleotide primers FPADG ( 5′-CTGGATCCATGCAAGCACCCGACCAAC-3′ ) and RPADG ( 5′-CGGGAATTCCTACCAACTCGTTATTGATAGAGCGATAA-3′ ), and then cloned into pGEX-4T-1 (GE Healthcare, Upsala, Sweden) flanked by Bam HI and Eco RI restriction sites.

    Techniques: Clone Assay, Expressing, Polymerase Chain Reaction, Amplification, Plasmid Preparation, Agarose Gel Electrophoresis, Sequencing, Binding Assay

    Western-blot analysis of the SCP2 domain Left panel: proteins from various tissues within the last larval instar were used. Lane 1, midgut; lane 2, fat body; lane 3, Malpighian tubules; lane 4, recombinant protein corresponding to the SCP2 domain of SCPx. The antibody against the SCP2 domain of S. littoralis SCPx protein was used. Right panel: Western-blot analysis of the recombinant S. littoralis SCPx protein using anti-(SCP2 domain) antibody. Lane 1, proteins from E. coli carrying the pGEX-2T vector without the SCPx cDNA; lane 2, proteins from E. coli carrying the pGEX-2T vector containing SCPx cDNA with induction of expression.

    Journal: Biochemical Journal

    Article Title: Characterization of a sterol carrier protein 2/3-oxoacyl-CoA thiolase from the cotton leafworm (Spodoptera littoralis): a lepidopteran mechanism closer to that in mammals than that in dipterans

    doi: 10.1042/BJ20040717

    Figure Lengend Snippet: Western-blot analysis of the SCP2 domain Left panel: proteins from various tissues within the last larval instar were used. Lane 1, midgut; lane 2, fat body; lane 3, Malpighian tubules; lane 4, recombinant protein corresponding to the SCP2 domain of SCPx. The antibody against the SCP2 domain of S. littoralis SCPx protein was used. Right panel: Western-blot analysis of the recombinant S. littoralis SCPx protein using anti-(SCP2 domain) antibody. Lane 1, proteins from E. coli carrying the pGEX-2T vector without the SCPx cDNA; lane 2, proteins from E. coli carrying the pGEX-2T vector containing SCPx cDNA with induction of expression.

    Article Snippet: The product was cut out with Bam HI and Eco RI from the plasmid and transferred into pGEX-2T (Amersham Biosciences).

    Techniques: Western Blot, Recombinant, Plasmid Preparation, Expressing

    The developmental profile of SCPx during the sixth instar ( A ) A 10 #x3BC;g portion of total RNA from midgut at various time points within the sixth larval instar, as indicated, was used. The blot was hybridized with a 32 P-labelled SCPx cDNA probe corresponding to the open reading frame. The same blot was stripped and reprobed with 18S rRNA probe. G, W and PP refer to the stages ‘gut purge’, ‘wandering’and ‘prepupa’ respectively that occur before pupation. ( B ) Western-blot analysis of SCPx protein. Top panel: 100 #x3BC;g of protein from midgut at various time points within the sixth larval instar, as indicated, was used. Bottom panel: Western-blot analysis of the recombinant S. littoralis SCPx protein. Lane 1, proteins from E. coli carrying the pGEX-2T vector without the SCPx cDNA; lane 2, proteins from E. coli carrying the pGEX-2T vector containing the SCPx cDNA without induction of expression; lane 3, proteins from E. coli carrying the pGEX-2T vector containing SCPx cDNA with induction of expression; lane 4, whole extract from C. elegans . The antibody against C. elegans P-44 was used to detect the SCPx protein.

    Journal: Biochemical Journal

    Article Title: Characterization of a sterol carrier protein 2/3-oxoacyl-CoA thiolase from the cotton leafworm (Spodoptera littoralis): a lepidopteran mechanism closer to that in mammals than that in dipterans

    doi: 10.1042/BJ20040717

    Figure Lengend Snippet: The developmental profile of SCPx during the sixth instar ( A ) A 10 #x3BC;g portion of total RNA from midgut at various time points within the sixth larval instar, as indicated, was used. The blot was hybridized with a 32 P-labelled SCPx cDNA probe corresponding to the open reading frame. The same blot was stripped and reprobed with 18S rRNA probe. G, W and PP refer to the stages ‘gut purge’, ‘wandering’and ‘prepupa’ respectively that occur before pupation. ( B ) Western-blot analysis of SCPx protein. Top panel: 100 #x3BC;g of protein from midgut at various time points within the sixth larval instar, as indicated, was used. Bottom panel: Western-blot analysis of the recombinant S. littoralis SCPx protein. Lane 1, proteins from E. coli carrying the pGEX-2T vector without the SCPx cDNA; lane 2, proteins from E. coli carrying the pGEX-2T vector containing the SCPx cDNA without induction of expression; lane 3, proteins from E. coli carrying the pGEX-2T vector containing SCPx cDNA with induction of expression; lane 4, whole extract from C. elegans . The antibody against C. elegans P-44 was used to detect the SCPx protein.

    Article Snippet: The product was cut out with Bam HI and Eco RI from the plasmid and transferred into pGEX-2T (Amersham Biosciences).

    Techniques: Western Blot, Recombinant, Plasmid Preparation, Expressing

    Schematic diagram of experimental approach. A PCR product containing 17 random codons was inserted into the BamHI site of pGEX-2TK producing various glutathione-S-transferase fusion proteins which were bound to glutathione-sepharose, and labeled with 32 P using protein kinase A. After washing and thrombin digestion, the labeled peptides were incubated with several different cell lines and assayed.

    Journal: PLoS ONE

    Article Title: Generation of Small 32P-Labeled Peptides as a Potential Approach to Colorectal Cancer Therapy

    doi: 10.1371/journal.pone.0002508

    Figure Lengend Snippet: Schematic diagram of experimental approach. A PCR product containing 17 random codons was inserted into the BamHI site of pGEX-2TK producing various glutathione-S-transferase fusion proteins which were bound to glutathione-sepharose, and labeled with 32 P using protein kinase A. After washing and thrombin digestion, the labeled peptides were incubated with several different cell lines and assayed.

    Article Snippet: Production of the recombinant 32 P-labeled peptides As described in , PCR generated products consisting of 17 random codons flanked by Bam HI sites were cloned into the Bam HI site of pGEX-2TK (GE Healthcare).

    Techniques: Polymerase Chain Reaction, Labeling, Incubation

    Establishment of monkey podoplanin-neutralizing antibodies (A) Purification of a monkey podoplanin (mkyPDPN) immunogen to establish a hybridoma secreting anti-PDPN monoclonal antibodies (mAbs). A mkyPDPN cDNA region encoding amino acids 76–89 (226–267 bp) was tandemly connected 21 times. The cDNA fragment was inserted into a pGEX-6P-3 vector, and a glutathione S-transferase (GST)-tagged mkyPDPN peptide (76–89 aa) produced by BL21 (DE3) E. coli was purified using glutathione sepharose. BALB/c mice were injected with the GST-tagged peptide, after which their splenocytes were fused with mouse myeloma P3U1 cells using polyethylene glycol. Hybridoma screening and antibody purification from ascites were performed. (B) CHO cells transfected with an empty vector (mock), wild-type monkey podoplanin (mkyPDPN-WT), wild-type human podoplanin (hPDPN-WT) were treated with PBS (closed areas) or antibodies (open areas), including anti-PDPN mAb D2-40, 1F6, 2F7, or 3F4 to measure PDPN expression levels. (C) GST-tagged recombinant mkyPDPN protein (WT) and its point mutants were expressed in E. coli . Cell lysates were electrophoresed and immunoblotted with antibodies to PDPN (1F6, 2F7, 3F4) or GST. The PLAG4 domain is indicated by red letters. (D) CHO/mock, CHO/mkyPDPN-WT, or CHO/hPDPN-WT cells were incubated with 100 μg/mL of control IgG1 or anti-PDPN antibodies 1F6, 2F7, or 3F4, followed by incubation with 1 μg/mL of hCLEC-2-(His) 10 (open areas: control IgG-treated samples; green areas: anti-PDPN mAb-treated samples). After washing, cells were further incubated with Alexa Flour 488-conjugated anti-penta-His second antibody. CLEC-2 binding was measured by flow cytometry. Gray areas indicate the fluorescence intensity of samples not treated with CLEC-2. (E) CHO/mkyPDPN-WT cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs followed by incubation with mouse platelet-rich plasma (PRP). The aggregation rate was estimated using an aggregometer. (F) PLAG3 domain-deleted mkyPDPN mutant cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs, followed by incubation with mouse PRP. The aggregation rate was estimated using an aggregometer.

    Journal: Oncotarget

    Article Title: A safety study of newly generated anti-podoplanin-neutralizing antibody in cynomolgus monkey (Macaca fascicularis)

    doi: 10.18632/oncotarget.26055

    Figure Lengend Snippet: Establishment of monkey podoplanin-neutralizing antibodies (A) Purification of a monkey podoplanin (mkyPDPN) immunogen to establish a hybridoma secreting anti-PDPN monoclonal antibodies (mAbs). A mkyPDPN cDNA region encoding amino acids 76–89 (226–267 bp) was tandemly connected 21 times. The cDNA fragment was inserted into a pGEX-6P-3 vector, and a glutathione S-transferase (GST)-tagged mkyPDPN peptide (76–89 aa) produced by BL21 (DE3) E. coli was purified using glutathione sepharose. BALB/c mice were injected with the GST-tagged peptide, after which their splenocytes were fused with mouse myeloma P3U1 cells using polyethylene glycol. Hybridoma screening and antibody purification from ascites were performed. (B) CHO cells transfected with an empty vector (mock), wild-type monkey podoplanin (mkyPDPN-WT), wild-type human podoplanin (hPDPN-WT) were treated with PBS (closed areas) or antibodies (open areas), including anti-PDPN mAb D2-40, 1F6, 2F7, or 3F4 to measure PDPN expression levels. (C) GST-tagged recombinant mkyPDPN protein (WT) and its point mutants were expressed in E. coli . Cell lysates were electrophoresed and immunoblotted with antibodies to PDPN (1F6, 2F7, 3F4) or GST. The PLAG4 domain is indicated by red letters. (D) CHO/mock, CHO/mkyPDPN-WT, or CHO/hPDPN-WT cells were incubated with 100 μg/mL of control IgG1 or anti-PDPN antibodies 1F6, 2F7, or 3F4, followed by incubation with 1 μg/mL of hCLEC-2-(His) 10 (open areas: control IgG-treated samples; green areas: anti-PDPN mAb-treated samples). After washing, cells were further incubated with Alexa Flour 488-conjugated anti-penta-His second antibody. CLEC-2 binding was measured by flow cytometry. Gray areas indicate the fluorescence intensity of samples not treated with CLEC-2. (E) CHO/mkyPDPN-WT cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs followed by incubation with mouse platelet-rich plasma (PRP). The aggregation rate was estimated using an aggregometer. (F) PLAG3 domain-deleted mkyPDPN mutant cells were incubated with 10 μg/mL of control IgG1, 1F6, 2F7, or 3F4 mAbs, followed by incubation with mouse PRP. The aggregation rate was estimated using an aggregometer.

    Article Snippet: The PCR products were subcloned by Topo TA cloning (Invitrogen, Carlsbad, CA, USA) and, after sequence confirmation, cloned into pcDNA3 (Life Technologies, Carlsbad, CA, USA) or pGEX-6P-3 (GE Healthcare, Buckinghamshire, UK) plasmids using EcoRI restriction sites, resulting in pcDNA3 monkey podoplanin or pGEX-6P-3 monkey podoplanin, respectively.

    Techniques: Purification, Plasmid Preparation, Produced, Mouse Assay, Injection, Antibody Purification, Transfection, Expressing, Recombinant, Incubation, Binding Assay, Flow Cytometry, Cytometry, Fluorescence, Mutagenesis

    Western blot analysis of Itih5 protein with the α-Itih5 antibody. (A) Characterization of the α-Itih5 antibody. Western blot analysis of protein extracts from E. coli strain BL21 producing the pGEX-3X/Itih5 recombinant protein. Lane 1: Protein extract without IPTG induction; lane 2: Protein extract with IPTG induction. The arrow indicates the molecular size of the specific signal for the GST-Itih5 protein (approximately 105 kDa). (B) Western blot characterization of the Itih5 isoforms produced in the female genital tract and the liver at E18.5. Arrows indicate the molecular sizes of the specific bands found in the embryonic female genital tract (105 and 50 kDa) and in the liver (73 kDa). (C) Size characterization of the Itih5 isoform produced in the uterus of pregnant and nonpregnant adult mice. The arrow indicates the molecular size of the specific band found in the uterus during gestation and in nonpregnant animals (73 kDa). E, embryonic day; NP, nonpregnant.

    Journal: Gene Expression

    Article Title: Involvement of ITIH5, a Candidate Gene for Congenital Uterovaginal Aplasia (Mayer-Rokitansky-Küster-Hauser Syndrome), in Female Genital Tract Development

    doi:

    Figure Lengend Snippet: Western blot analysis of Itih5 protein with the α-Itih5 antibody. (A) Characterization of the α-Itih5 antibody. Western blot analysis of protein extracts from E. coli strain BL21 producing the pGEX-3X/Itih5 recombinant protein. Lane 1: Protein extract without IPTG induction; lane 2: Protein extract with IPTG induction. The arrow indicates the molecular size of the specific signal for the GST-Itih5 protein (approximately 105 kDa). (B) Western blot characterization of the Itih5 isoforms produced in the female genital tract and the liver at E18.5. Arrows indicate the molecular sizes of the specific bands found in the embryonic female genital tract (105 and 50 kDa) and in the liver (73 kDa). (C) Size characterization of the Itih5 isoform produced in the uterus of pregnant and nonpregnant adult mice. The arrow indicates the molecular size of the specific band found in the uterus during gestation and in nonpregnant animals (73 kDa). E, embryonic day; NP, nonpregnant.

    Article Snippet: A cDNA fragment corresponding to amino acids 219–952 of the Itih5 protein was inserted into pGEX-3X (GE Healthcare, Saclay, France) and the resulting recombinant plasmid was used to produce a GST-fusion protein, GST-Itih5.

    Techniques: Western Blot, Recombinant, Produced, Mouse Assay

    Concept of the asymmetric directional T-vector. ( A ) Construction of pGEX-4T3-PRESAT and direct cloning of PCR product in pGEX-4T3-PRESAT. ( B ) The schematic representation of the ORF selection method using potential restriction enzyme site. The figure illustrates the case in which NcoI is chosen as the second restriction enzyme for selection. The rear PCR primer is designed with 5′-GG at the 5′ end, so that only the ligated plasmid with insert in the reverse orientation will have the NcoI site at the TA-cloning position. For NdeI selection, the rear primer with 5′-ATG is used instead of the 5′-GG primer.

    Journal: Protein Science : A Publication of the Protein Society

    Article Title: The PRESAT-vector: Asymmetric T-vector for high-throughput screening of soluble protein domains for structural proteomics

    doi: 10.1110/ps.03439004

    Figure Lengend Snippet: Concept of the asymmetric directional T-vector. ( A ) Construction of pGEX-4T3-PRESAT and direct cloning of PCR product in pGEX-4T3-PRESAT. ( B ) The schematic representation of the ORF selection method using potential restriction enzyme site. The figure illustrates the case in which NcoI is chosen as the second restriction enzyme for selection. The rear PCR primer is designed with 5′-GG at the 5′ end, so that only the ligated plasmid with insert in the reverse orientation will have the NcoI site at the TA-cloning position. For NdeI selection, the rear primer with 5′-ATG is used instead of the 5′-GG primer.

    Article Snippet: Oligonucleotide primers were obtained from Hokkaido System Science. pGEM-T (Promega), pGEX-4T3 (Amersham Bioscience), and pET32a (Novagen), respectively, were purchased.

    Techniques: Plasmid Preparation, Clone Assay, Polymerase Chain Reaction, Selection, TA Cloning

    Concept of HTS for the soluble GST-fusion proteins from protein expression libraries. (i) Various PCR primers were designed and used to amplify a series of partial cDNA fragments of the protein of interest, which may contain a putative domain. (ii) The fragments are inserted into pGEX-4T3-PRESAT vector, selected for ORF orientation by restriction cleavage, and transformed into E. coli for colony isolation. (iii) Deep 96-well format plates are used to culture individual clones to allow expression of the GST-fusion proteins. (iv) The clone displaying the highest CDNB assay signal will be selected for further analysis.

    Journal: Protein Science : A Publication of the Protein Society

    Article Title: The PRESAT-vector: Asymmetric T-vector for high-throughput screening of soluble protein domains for structural proteomics

    doi: 10.1110/ps.03439004

    Figure Lengend Snippet: Concept of HTS for the soluble GST-fusion proteins from protein expression libraries. (i) Various PCR primers were designed and used to amplify a series of partial cDNA fragments of the protein of interest, which may contain a putative domain. (ii) The fragments are inserted into pGEX-4T3-PRESAT vector, selected for ORF orientation by restriction cleavage, and transformed into E. coli for colony isolation. (iii) Deep 96-well format plates are used to culture individual clones to allow expression of the GST-fusion proteins. (iv) The clone displaying the highest CDNB assay signal will be selected for further analysis.

    Article Snippet: Oligonucleotide primers were obtained from Hokkaido System Science. pGEM-T (Promega), pGEX-4T3 (Amersham Bioscience), and pET32a (Novagen), respectively, were purchased.

    Techniques: Expressing, Polymerase Chain Reaction, Plasmid Preparation, Transformation Assay, Isolation, Clone Assay

    The presence of 14-antigens in patients with anti-GST antibodies To purify the SEREX-identified proteins, the insertion sequences of the 14 pBluescript plasmids were ligated in-frame into GST-tagged expression vectors. We confirmed by sequence analysis that the recombinant pGEX-4T-3 plasmids were properly recombined and GST-tagged recombinant proteins were affinity-purified using glutathione-Sepharose. To confirm the recombinant proteins to be the GST-tagged one that react with autologous plasma, the proteins were lysed in a SDS sample buffer, incubated at 100°C for 3 min, Affinity-purified GST-tagged antigens were separated on 11% SDS-polyacrylamide gels followed by Western blot using anti-GST antibody. All samples were examined simultaneously, at the same time on the same membrane.

    Journal: Oncotarget

    Article Title: Identification of specific and common diagnostic antibody markers for gastrointestinal cancers by SEREX screening using testis cDNA phage library

    doi: 10.18632/oncotarget.24963

    Figure Lengend Snippet: The presence of 14-antigens in patients with anti-GST antibodies To purify the SEREX-identified proteins, the insertion sequences of the 14 pBluescript plasmids were ligated in-frame into GST-tagged expression vectors. We confirmed by sequence analysis that the recombinant pGEX-4T-3 plasmids were properly recombined and GST-tagged recombinant proteins were affinity-purified using glutathione-Sepharose. To confirm the recombinant proteins to be the GST-tagged one that react with autologous plasma, the proteins were lysed in a SDS sample buffer, incubated at 100°C for 3 min, Affinity-purified GST-tagged antigens were separated on 11% SDS-polyacrylamide gels followed by Western blot using anti-GST antibody. All samples were examined simultaneously, at the same time on the same membrane.

    Article Snippet: The expression plasmids of glutathione S- transferase (GST)-fused proteins were constructed by recombining the cDNA sequences into pGEX-4T-3 (GE Healthcare Life Sciences, Pittsburgh, PA).

    Techniques: Expressing, Sequencing, Recombinant, Affinity Purification, Incubation, Western Blot

    HCV genome and recombinant proteins. (A) The HCV genome contains a single major open reading frame (ORF) flanked by untranslated regions (UTRs). The 10 proteins encoded within the main ORF are indicated by alternated shading. (B) Genomic fragments of HCV were cloned into the prokaryotic expression vectors, pET32a(+), pQE30, or pGEX-4T-2, in-frame downstream of the 6-His-tag or glutathione S-transferase (GST)-tag coding sequence.

    Journal: International Journal of Molecular Medicine

    Article Title: A novel double antibody sandwich-lateral flow immunoassay for the rapid and simple detection of hepatitis C virus

    doi: 10.3892/ijmm.2012.1121

    Figure Lengend Snippet: HCV genome and recombinant proteins. (A) The HCV genome contains a single major open reading frame (ORF) flanked by untranslated regions (UTRs). The 10 proteins encoded within the main ORF are indicated by alternated shading. (B) Genomic fragments of HCV were cloned into the prokaryotic expression vectors, pET32a(+), pQE30, or pGEX-4T-2, in-frame downstream of the 6-His-tag or glutathione S-transferase (GST)-tag coding sequence.

    Article Snippet: Construction of expression plasmids For the expression of HCV proteins in E. coli , corresponding coding sequences were cloned into the histidine fusion expression vectors, pET32a(+) and pQE30, or the GST-tag expression vector, pGEX-4T-2, as shown in and .

    Techniques: Recombinant, Clone Assay, Expressing, Sequencing