tbwee1 Search Results


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  • 99
    Thermo Fisher ecori
    Ecori, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 12201 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore synthetic peptide poly
    Synthetic Peptide Poly, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher pfastbactm
    Pfastbactm, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Thermo Fisher destination vector pdest17
    Amplification and purification of TbWee1-6xHis fusion protein to produce anti-TbWee1 polyclonal antibodies. (A) SDS–PAGE analysis of lysates obtained from Escherichia coli IPTG.induced (I) and non-induced (NI) cell cultures transformed with <t>pDEST17-TbWee1.</t> Upper panel: Coomassie Brilliant Blue staining. Lower panel: Western-blot analysis with anti-histidine tag antibody (1:5000). The arrow indicates the position of the ~70 kDa recombinant protein. (B) Purification steps of rTbwee1-6xHis fusion protein were monitored by 12 % SDS-PAGE analysis. The lysate was loaded onto a Ni-agarose column. Flow-through (F), washes (W, 1–3), and eluted fractions (250 mM imidazol, E 1–3) were analyzed by 12% SDS-PAGE stained with Coomassie Brilliant Blue. (C) Western blot analysis of rTbWee1 expression with polyclonal anti-TbWee1 antibody. Protein extracts of the T. brucei bloodstream form (lane 1), the T. brucei procyclic form (lane 2) and the recombinant TbWee1-6xHis (lane 3) were separated by 10 % SDS-PAGE and electroblotted on nitrocellulose membrane. Blots were incubated with anti-TbWee1 polyclonal antibody (1:1000) and revealed by chemiluminescence.
    Destination Vector Pdest17, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 120 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher gateway entry vector pentr 2b
    Amplification and purification of TbWee1-6xHis fusion protein to produce anti-TbWee1 polyclonal antibodies. (A) SDS–PAGE analysis of lysates obtained from Escherichia coli IPTG.induced (I) and non-induced (NI) cell cultures transformed with <t>pDEST17-TbWee1.</t> Upper panel: Coomassie Brilliant Blue staining. Lower panel: Western-blot analysis with anti-histidine tag antibody (1:5000). The arrow indicates the position of the ~70 kDa recombinant protein. (B) Purification steps of rTbwee1-6xHis fusion protein were monitored by 12 % SDS-PAGE analysis. The lysate was loaded onto a Ni-agarose column. Flow-through (F), washes (W, 1–3), and eluted fractions (250 mM imidazol, E 1–3) were analyzed by 12% SDS-PAGE stained with Coomassie Brilliant Blue. (C) Western blot analysis of rTbWee1 expression with polyclonal anti-TbWee1 antibody. Protein extracts of the T. brucei bloodstream form (lane 1), the T. brucei procyclic form (lane 2) and the recombinant TbWee1-6xHis (lane 3) were separated by 10 % SDS-PAGE and electroblotted on nitrocellulose membrane. Blots were incubated with anti-TbWee1 polyclonal antibody (1:1000) and revealed by chemiluminescence.
    Gateway Entry Vector Pentr 2b, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore histones
    Amplification and purification of TbWee1-6xHis fusion protein to produce anti-TbWee1 polyclonal antibodies. (A) SDS–PAGE analysis of lysates obtained from Escherichia coli IPTG.induced (I) and non-induced (NI) cell cultures transformed with <t>pDEST17-TbWee1.</t> Upper panel: Coomassie Brilliant Blue staining. Lower panel: Western-blot analysis with anti-histidine tag antibody (1:5000). The arrow indicates the position of the ~70 kDa recombinant protein. (B) Purification steps of rTbwee1-6xHis fusion protein were monitored by 12 % SDS-PAGE analysis. The lysate was loaded onto a Ni-agarose column. Flow-through (F), washes (W, 1–3), and eluted fractions (250 mM imidazol, E 1–3) were analyzed by 12% SDS-PAGE stained with Coomassie Brilliant Blue. (C) Western blot analysis of rTbWee1 expression with polyclonal anti-TbWee1 antibody. Protein extracts of the T. brucei bloodstream form (lane 1), the T. brucei procyclic form (lane 2) and the recombinant TbWee1-6xHis (lane 3) were separated by 10 % SDS-PAGE and electroblotted on nitrocellulose membrane. Blots were incubated with anti-TbWee1 polyclonal antibody (1:1000) and revealed by chemiluminescence.
    Histones, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 289 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher gateway lr clonase ii enzyme mix kit
    Amplification and purification of TbWee1-6xHis fusion protein to produce anti-TbWee1 polyclonal antibodies. (A) SDS–PAGE analysis of lysates obtained from Escherichia coli IPTG.induced (I) and non-induced (NI) cell cultures transformed with <t>pDEST17-TbWee1.</t> Upper panel: Coomassie Brilliant Blue staining. Lower panel: Western-blot analysis with anti-histidine tag antibody (1:5000). The arrow indicates the position of the ~70 kDa recombinant protein. (B) Purification steps of rTbwee1-6xHis fusion protein were monitored by 12 % SDS-PAGE analysis. The lysate was loaded onto a Ni-agarose column. Flow-through (F), washes (W, 1–3), and eluted fractions (250 mM imidazol, E 1–3) were analyzed by 12% SDS-PAGE stained with Coomassie Brilliant Blue. (C) Western blot analysis of rTbWee1 expression with polyclonal anti-TbWee1 antibody. Protein extracts of the T. brucei bloodstream form (lane 1), the T. brucei procyclic form (lane 2) and the recombinant TbWee1-6xHis (lane 3) were separated by 10 % SDS-PAGE and electroblotted on nitrocellulose membrane. Blots were incubated with anti-TbWee1 polyclonal antibody (1:1000) and revealed by chemiluminescence.
    Gateway Lr Clonase Ii Enzyme Mix Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 96 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Stratagene t brucei
    Effects of TbWee1 knockdown on the procyclic form of T. <t>brucei</t> cells. (A) Cells of strain 29-13 harboring the TbWee1-RNAi construct were incubated in culture medium with (+ Tet) or without (-Tet) 2.5 µg/ml tetracycline at 28°C. The cell growth rate was monitored daily, and the cell number was plotted in a logarithmic scale. The insets show the intracellular mRNA level after 3 days of RNAi as monitored by Northern blot. RNAr was used as loading control. Western blot of extracts of induced and non-induced cells were analyzed with anti-TbWee1 antibody (Right inset). (B) Time course of RNAi-induced T. brucei procyclic-form. Cells were stained with propidium iodide and subjected to FACS analysis to measure <t>DNA</t> content. The percentages of cells in G1, S and G2/M phases were determined with the ModFitLT software and plotted on the right panel.
    T Brucei, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Stratagene pfu polymerase
    Effects of TbWee1 knockdown on the procyclic form of T. <t>brucei</t> cells. (A) Cells of strain 29-13 harboring the TbWee1-RNAi construct were incubated in culture medium with (+ Tet) or without (-Tet) 2.5 µg/ml tetracycline at 28°C. The cell growth rate was monitored daily, and the cell number was plotted in a logarithmic scale. The insets show the intracellular mRNA level after 3 days of RNAi as monitored by Northern blot. RNAr was used as loading control. Western blot of extracts of induced and non-induced cells were analyzed with anti-TbWee1 antibody (Right inset). (B) Time course of RNAi-induced T. brucei procyclic-form. Cells were stained with propidium iodide and subjected to FACS analysis to measure <t>DNA</t> content. The percentages of cells in G1, S and G2/M phases were determined with the ModFitLT software and plotted on the right panel.
    Pfu Polymerase, supplied by Stratagene, used in various techniques. Bioz Stars score: 93/100, based on 10176 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher spodoptera frugiperda sf 9 insect cells
    Effects of TbWee1 knockdown on the procyclic form of T. <t>brucei</t> cells. (A) Cells of strain 29-13 harboring the TbWee1-RNAi construct were incubated in culture medium with (+ Tet) or without (-Tet) 2.5 µg/ml tetracycline at 28°C. The cell growth rate was monitored daily, and the cell number was plotted in a logarithmic scale. The insets show the intracellular mRNA level after 3 days of RNAi as monitored by Northern blot. RNAr was used as loading control. Western blot of extracts of induced and non-induced cells were analyzed with anti-TbWee1 antibody (Right inset). (B) Time course of RNAi-induced T. brucei procyclic-form. Cells were stained with propidium iodide and subjected to FACS analysis to measure <t>DNA</t> content. The percentages of cells in G1, S and G2/M phases were determined with the ModFitLT software and plotted on the right panel.
    Spodoptera Frugiperda Sf 9 Insect Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 91 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Macrogen pcr induced mutations
    Effects of TbWee1 knockdown on the procyclic form of T. <t>brucei</t> cells. (A) Cells of strain 29-13 harboring the TbWee1-RNAi construct were incubated in culture medium with (+ Tet) or without (-Tet) 2.5 µg/ml tetracycline at 28°C. The cell growth rate was monitored daily, and the cell number was plotted in a logarithmic scale. The insets show the intracellular mRNA level after 3 days of RNAi as monitored by Northern blot. RNAr was used as loading control. Western blot of extracts of induced and non-induced cells were analyzed with anti-TbWee1 antibody (Right inset). (B) Time course of RNAi-induced T. brucei procyclic-form. Cells were stained with propidium iodide and subjected to FACS analysis to measure <t>DNA</t> content. The percentages of cells in G1, S and G2/M phases were determined with the ModFitLT software and plotted on the right panel.
    Pcr Induced Mutations, supplied by Macrogen, used in various techniques. Bioz Stars score: 88/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Promega prime a gene labeling system
    Effects of TbWee1 knockdown on the procyclic form of T. <t>brucei</t> cells. (A) Cells of strain 29-13 harboring the TbWee1-RNAi construct were incubated in culture medium with (+ Tet) or without (-Tet) 2.5 µg/ml tetracycline at 28°C. The cell growth rate was monitored daily, and the cell number was plotted in a logarithmic scale. The insets show the intracellular mRNA level after 3 days of RNAi as monitored by Northern blot. RNAr was used as loading control. Western blot of extracts of induced and non-induced cells were analyzed with anti-TbWee1 antibody (Right inset). (B) Time course of RNAi-induced T. brucei procyclic-form. Cells were stained with propidium iodide and subjected to FACS analysis to measure <t>DNA</t> content. The percentages of cells in G1, S and G2/M phases were determined with the ModFitLT software and plotted on the right panel.
    Prime A Gene Labeling System, supplied by Promega, used in various techniques. Bioz Stars score: 97/100, based on 2049 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Millipore rtbwee1
    Autophosphorylation of recombinant TbWee1-6xHis fusion protein. Phosphorylation reactions were performed in the absence or presence of 40 µg of <t>rTbWee1-6xHis</t> expressed in the baculovirus system, in a mixture containing 5 µCi [γ-32P]-ATP (6000 Ci/mmol, NEN) for 10 min at 30°C. Reaction products were resolved by 12 % SDS-PAGE and visualized by autoradiography. Membranes were revealed with anti-histidine tag antibody (1:5000) and anti-phosphotyrosine antibodies (1:1000).
    Rtbwee1, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen nickel affinity chromatography
    Autophosphorylation of recombinant TbWee1-6xHis fusion protein. Phosphorylation reactions were performed in the absence or presence of 40 µg of <t>rTbWee1-6xHis</t> expressed in the baculovirus system, in a mixture containing 5 µCi [γ-32P]-ATP (6000 Ci/mmol, NEN) for 10 min at 30°C. Reaction products were resolved by 12 % SDS-PAGE and visualized by autoradiography. Membranes were revealed with anti-histidine tag antibody (1:5000) and anti-phosphotyrosine antibodies (1:1000).
    Nickel Affinity Chromatography, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 2545 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher phusion dna polymerase
    Autophosphorylation of recombinant TbWee1-6xHis fusion protein. Phosphorylation reactions were performed in the absence or presence of 40 µg of <t>rTbWee1-6xHis</t> expressed in the baculovirus system, in a mixture containing 5 µCi [γ-32P]-ATP (6000 Ci/mmol, NEN) for 10 min at 30°C. Reaction products were resolved by 12 % SDS-PAGE and visualized by autoradiography. Membranes were revealed with anti-histidine tag antibody (1:5000) and anti-phosphotyrosine antibodies (1:1000).
    Phusion Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10742 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher trizol reagent
    Autophosphorylation of recombinant TbWee1-6xHis fusion protein. Phosphorylation reactions were performed in the absence or presence of 40 µg of <t>rTbWee1-6xHis</t> expressed in the baculovirus system, in a mixture containing 5 µCi [γ-32P]-ATP (6000 Ci/mmol, NEN) for 10 min at 30°C. Reaction products were resolved by 12 % SDS-PAGE and visualized by autoradiography. Membranes were revealed with anti-histidine tag antibody (1:5000) and anti-phosphotyrosine antibodies (1:1000).
    Trizol Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 604295 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Amplification and purification of TbWee1-6xHis fusion protein to produce anti-TbWee1 polyclonal antibodies. (A) SDS–PAGE analysis of lysates obtained from Escherichia coli IPTG.induced (I) and non-induced (NI) cell cultures transformed with pDEST17-TbWee1. Upper panel: Coomassie Brilliant Blue staining. Lower panel: Western-blot analysis with anti-histidine tag antibody (1:5000). The arrow indicates the position of the ~70 kDa recombinant protein. (B) Purification steps of rTbwee1-6xHis fusion protein were monitored by 12 % SDS-PAGE analysis. The lysate was loaded onto a Ni-agarose column. Flow-through (F), washes (W, 1–3), and eluted fractions (250 mM imidazol, E 1–3) were analyzed by 12% SDS-PAGE stained with Coomassie Brilliant Blue. (C) Western blot analysis of rTbWee1 expression with polyclonal anti-TbWee1 antibody. Protein extracts of the T. brucei bloodstream form (lane 1), the T. brucei procyclic form (lane 2) and the recombinant TbWee1-6xHis (lane 3) were separated by 10 % SDS-PAGE and electroblotted on nitrocellulose membrane. Blots were incubated with anti-TbWee1 polyclonal antibody (1:1000) and revealed by chemiluminescence.

    Journal: PLoS ONE

    Article Title: Identification of a Wee1-Like Kinase Gene Essential for Procyclic Trypanosoma brucei Survival

    doi: 10.1371/journal.pone.0079364

    Figure Lengend Snippet: Amplification and purification of TbWee1-6xHis fusion protein to produce anti-TbWee1 polyclonal antibodies. (A) SDS–PAGE analysis of lysates obtained from Escherichia coli IPTG.induced (I) and non-induced (NI) cell cultures transformed with pDEST17-TbWee1. Upper panel: Coomassie Brilliant Blue staining. Lower panel: Western-blot analysis with anti-histidine tag antibody (1:5000). The arrow indicates the position of the ~70 kDa recombinant protein. (B) Purification steps of rTbwee1-6xHis fusion protein were monitored by 12 % SDS-PAGE analysis. The lysate was loaded onto a Ni-agarose column. Flow-through (F), washes (W, 1–3), and eluted fractions (250 mM imidazol, E 1–3) were analyzed by 12% SDS-PAGE stained with Coomassie Brilliant Blue. (C) Western blot analysis of rTbWee1 expression with polyclonal anti-TbWee1 antibody. Protein extracts of the T. brucei bloodstream form (lane 1), the T. brucei procyclic form (lane 2) and the recombinant TbWee1-6xHis (lane 3) were separated by 10 % SDS-PAGE and electroblotted on nitrocellulose membrane. Blots were incubated with anti-TbWee1 polyclonal antibody (1:1000) and revealed by chemiluminescence.

    Article Snippet: The Gateway LR Clonase II Enzyme Mix kit (Invitrogen) was used to insert the TbWee1 coding sequence into the destination vector pDEST17 (Invitrogen) to generate the recombinant protein with an N-terminal histidine tag.

    Techniques: Amplification, Purification, SDS Page, Transformation Assay, Staining, Western Blot, Recombinant, Flow Cytometry, Expressing, Incubation

    Effects of TbWee1 knockdown on the procyclic form of T. brucei cells. (A) Cells of strain 29-13 harboring the TbWee1-RNAi construct were incubated in culture medium with (+ Tet) or without (-Tet) 2.5 µg/ml tetracycline at 28°C. The cell growth rate was monitored daily, and the cell number was plotted in a logarithmic scale. The insets show the intracellular mRNA level after 3 days of RNAi as monitored by Northern blot. RNAr was used as loading control. Western blot of extracts of induced and non-induced cells were analyzed with anti-TbWee1 antibody (Right inset). (B) Time course of RNAi-induced T. brucei procyclic-form. Cells were stained with propidium iodide and subjected to FACS analysis to measure DNA content. The percentages of cells in G1, S and G2/M phases were determined with the ModFitLT software and plotted on the right panel.

    Journal: PLoS ONE

    Article Title: Identification of a Wee1-Like Kinase Gene Essential for Procyclic Trypanosoma brucei Survival

    doi: 10.1371/journal.pone.0079364

    Figure Lengend Snippet: Effects of TbWee1 knockdown on the procyclic form of T. brucei cells. (A) Cells of strain 29-13 harboring the TbWee1-RNAi construct were incubated in culture medium with (+ Tet) or without (-Tet) 2.5 µg/ml tetracycline at 28°C. The cell growth rate was monitored daily, and the cell number was plotted in a logarithmic scale. The insets show the intracellular mRNA level after 3 days of RNAi as monitored by Northern blot. RNAr was used as loading control. Western blot of extracts of induced and non-induced cells were analyzed with anti-TbWee1 antibody (Right inset). (B) Time course of RNAi-induced T. brucei procyclic-form. Cells were stained with propidium iodide and subjected to FACS analysis to measure DNA content. The percentages of cells in G1, S and G2/M phases were determined with the ModFitLT software and plotted on the right panel.

    Article Snippet: It was amplified from genomic DNA obtained from T. brucei by PCR using the oligonucleotides 5´- GAATTC ATGTTGGCGCCTAAAGGGG-3´ and 5´-CG GATATC CTAAAATTTTGCACTATCTCC-3´ (restriction sites corresponding to EcoRI and EcoRV are underlined) using Pfu polymerase (Stratagene, La Jolla, USA).

    Techniques: Construct, Incubation, Northern Blot, Western Blot, Staining, FACS, Software

    Autophosphorylation of recombinant TbWee1-6xHis fusion protein. Phosphorylation reactions were performed in the absence or presence of 40 µg of rTbWee1-6xHis expressed in the baculovirus system, in a mixture containing 5 µCi [γ-32P]-ATP (6000 Ci/mmol, NEN) for 10 min at 30°C. Reaction products were resolved by 12 % SDS-PAGE and visualized by autoradiography. Membranes were revealed with anti-histidine tag antibody (1:5000) and anti-phosphotyrosine antibodies (1:1000).

    Journal: PLoS ONE

    Article Title: Identification of a Wee1-Like Kinase Gene Essential for Procyclic Trypanosoma brucei Survival

    doi: 10.1371/journal.pone.0079364

    Figure Lengend Snippet: Autophosphorylation of recombinant TbWee1-6xHis fusion protein. Phosphorylation reactions were performed in the absence or presence of 40 µg of rTbWee1-6xHis expressed in the baculovirus system, in a mixture containing 5 µCi [γ-32P]-ATP (6000 Ci/mmol, NEN) for 10 min at 30°C. Reaction products were resolved by 12 % SDS-PAGE and visualized by autoradiography. Membranes were revealed with anti-histidine tag antibody (1:5000) and anti-phosphotyrosine antibodies (1:1000).

    Article Snippet: These assays were performed with 40 µg of rTbwee1 either with 0.2 mg/ml poly-(glu:tyr) (Sigma-Aldrich, St. Louis, Missouri, USA), a general substrate for tyrosine protein kinase, and analyzed with P81 phosphocellulose papers [44], or with 0.5 mg ml-1 of histone H1 as previously reported [39].

    Techniques: Recombinant, SDS Page, Autoradiography

    Amplification and purification of TbWee1-6xHis fusion protein to produce anti-TbWee1 polyclonal antibodies. (A) SDS–PAGE analysis of lysates obtained from Escherichia coli IPTG.induced (I) and non-induced (NI) cell cultures transformed with pDEST17-TbWee1. Upper panel: Coomassie Brilliant Blue staining. Lower panel: Western-blot analysis with anti-histidine tag antibody (1:5000). The arrow indicates the position of the ~70 kDa recombinant protein. (B) Purification steps of rTbwee1-6xHis fusion protein were monitored by 12 % SDS-PAGE analysis. The lysate was loaded onto a Ni-agarose column. Flow-through (F), washes (W, 1–3), and eluted fractions (250 mM imidazol, E 1–3) were analyzed by 12% SDS-PAGE stained with Coomassie Brilliant Blue. (C) Western blot analysis of rTbWee1 expression with polyclonal anti-TbWee1 antibody. Protein extracts of the T. brucei bloodstream form (lane 1), the T. brucei procyclic form (lane 2) and the recombinant TbWee1-6xHis (lane 3) were separated by 10 % SDS-PAGE and electroblotted on nitrocellulose membrane. Blots were incubated with anti-TbWee1 polyclonal antibody (1:1000) and revealed by chemiluminescence.

    Journal: PLoS ONE

    Article Title: Identification of a Wee1-Like Kinase Gene Essential for Procyclic Trypanosoma brucei Survival

    doi: 10.1371/journal.pone.0079364

    Figure Lengend Snippet: Amplification and purification of TbWee1-6xHis fusion protein to produce anti-TbWee1 polyclonal antibodies. (A) SDS–PAGE analysis of lysates obtained from Escherichia coli IPTG.induced (I) and non-induced (NI) cell cultures transformed with pDEST17-TbWee1. Upper panel: Coomassie Brilliant Blue staining. Lower panel: Western-blot analysis with anti-histidine tag antibody (1:5000). The arrow indicates the position of the ~70 kDa recombinant protein. (B) Purification steps of rTbwee1-6xHis fusion protein were monitored by 12 % SDS-PAGE analysis. The lysate was loaded onto a Ni-agarose column. Flow-through (F), washes (W, 1–3), and eluted fractions (250 mM imidazol, E 1–3) were analyzed by 12% SDS-PAGE stained with Coomassie Brilliant Blue. (C) Western blot analysis of rTbWee1 expression with polyclonal anti-TbWee1 antibody. Protein extracts of the T. brucei bloodstream form (lane 1), the T. brucei procyclic form (lane 2) and the recombinant TbWee1-6xHis (lane 3) were separated by 10 % SDS-PAGE and electroblotted on nitrocellulose membrane. Blots were incubated with anti-TbWee1 polyclonal antibody (1:1000) and revealed by chemiluminescence.

    Article Snippet: These assays were performed with 40 µg of rTbwee1 either with 0.2 mg/ml poly-(glu:tyr) (Sigma-Aldrich, St. Louis, Missouri, USA), a general substrate for tyrosine protein kinase, and analyzed with P81 phosphocellulose papers [44], or with 0.5 mg ml-1 of histone H1 as previously reported [39].

    Techniques: Amplification, Purification, SDS Page, Transformation Assay, Staining, Western Blot, Recombinant, Flow Cytometry, Expressing, Incubation