bl21 a1 bacteria  (Thermo Fisher)


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    Structured Review

    Thermo Fisher bl21 a1 bacteria
    Bl21 A1 Bacteria, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 a1 bacteria/product/Thermo Fisher
    Average 79 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    bl21 a1 bacteria - by Bioz Stars, 2020-02
    79/100 stars

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    Centrifugation:

    Article Title: PAK4: a pluripotent kinase that regulates prostate cancer cell adhesion
    Article Snippet: GST proteins were purified from BL21-A1 bacteria (Invitrogen) as previously described ( ). .. The pre-cleared lysates were incubated with the GST-fusion-protein beads for 3 hours at 4°C, collected by centrifugation, washed three times with wash buffer (20 mM Tris-HCl, pH 7.6, 300 mM NaCl, 2 mM EDTA, 1% Triton X-100, 10% glycerol, 50 mM NaF, 1 mM Na3 VO4 and protease-inhibitor cocktail) and resuspended in 2× SDS loading buffer.

    Article Title: A PAK6-IQGAP1 complex promotes disassembly of cell-cell adhesions
    Article Snippet: GST pulldown GST proteins were purified from BL21-A1 bacteria (Invitrogen) as previously described [ ]. .. Lysates were pre-cleared by incubation with GST-coupled Glutathione Sepharose 4 Fast Flow beads (Amersham) for 1 h at 4 °C, then incubated with the GST fusion protein beads for 2 h at 4 °C, collected by centrifugation, washed three times with lysis buffer, and resuspended in 2 × SDS loading buffer.

    Protease Inhibitor:

    Article Title: PAK4: a pluripotent kinase that regulates prostate cancer cell adhesion
    Article Snippet: GST proteins were purified from BL21-A1 bacteria (Invitrogen) as previously described ( ). .. HeLa cells were lysed in 20 mM Tris-HCl, pH 7.6, 150 mM NaCl, 2 mM EDTA, 0.1% Triton X-100, 50 mM NaF, 1 mM Na3 VO4 and protease-inhibitor cocktail (Roche).

    Flow Cytometry:

    Article Title: PAK4: a pluripotent kinase that regulates prostate cancer cell adhesion
    Article Snippet: GST proteins were purified from BL21-A1 bacteria (Invitrogen) as previously described ( ). .. Lysates were then pre-cleared by incubation with GST-coupled Glutathione Sepharose 4 Fast Flow beads (Amersham) for 1 hour at 4°C.

    Article Title: A PAK6-IQGAP1 complex promotes disassembly of cell-cell adhesions
    Article Snippet: GST pulldown GST proteins were purified from BL21-A1 bacteria (Invitrogen) as previously described [ ]. .. Lysates were pre-cleared by incubation with GST-coupled Glutathione Sepharose 4 Fast Flow beads (Amersham) for 1 h at 4 °C, then incubated with the GST fusion protein beads for 2 h at 4 °C, collected by centrifugation, washed three times with lysis buffer, and resuspended in 2 × SDS loading buffer.

    Purification:

    Article Title: PAK4: a pluripotent kinase that regulates prostate cancer cell adhesion
    Article Snippet: .. GST proteins were purified from BL21-A1 bacteria (Invitrogen) as previously described ( ). .. HeLa cells were lysed in 20 mM Tris-HCl, pH 7.6, 150 mM NaCl, 2 mM EDTA, 0.1% Triton X-100, 50 mM NaF, 1 mM Na3 VO4 and protease-inhibitor cocktail (Roche).

    Article Title: A PAK6-IQGAP1 complex promotes disassembly of cell-cell adhesions
    Article Snippet: .. GST pulldown GST proteins were purified from BL21-A1 bacteria (Invitrogen) as previously described [ ]. .. Cells were lysed in NP-40 lysis buffer (0.5 % NP-40, 50 mM Tris–HCl, pH 7.6, 30 mM sodium pyrophosphate, 150 mM NaCl, 0.1 mM EDTA, 50 mM NaF, 1 mM Na3 VO4 , 1 mM PMSF, 10 μg/ml leupeptin, 1 μg/ml aprotinin, and 1 mM DTT).

    Incubation:

    Article Title: PAK4: a pluripotent kinase that regulates prostate cancer cell adhesion
    Article Snippet: GST proteins were purified from BL21-A1 bacteria (Invitrogen) as previously described ( ). .. Lysates were then pre-cleared by incubation with GST-coupled Glutathione Sepharose 4 Fast Flow beads (Amersham) for 1 hour at 4°C.

    Article Title: A PAK6-IQGAP1 complex promotes disassembly of cell-cell adhesions
    Article Snippet: GST pulldown GST proteins were purified from BL21-A1 bacteria (Invitrogen) as previously described [ ]. .. Lysates were pre-cleared by incubation with GST-coupled Glutathione Sepharose 4 Fast Flow beads (Amersham) for 1 h at 4 °C, then incubated with the GST fusion protein beads for 2 h at 4 °C, collected by centrifugation, washed three times with lysis buffer, and resuspended in 2 × SDS loading buffer.

    Lysis:

    Article Title: A PAK6-IQGAP1 complex promotes disassembly of cell-cell adhesions
    Article Snippet: GST pulldown GST proteins were purified from BL21-A1 bacteria (Invitrogen) as previously described [ ]. .. Cells were lysed in NP-40 lysis buffer (0.5 % NP-40, 50 mM Tris–HCl, pH 7.6, 30 mM sodium pyrophosphate, 150 mM NaCl, 0.1 mM EDTA, 50 mM NaF, 1 mM Na3 VO4 , 1 mM PMSF, 10 μg/ml leupeptin, 1 μg/ml aprotinin, and 1 mM DTT).

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  • 99
    Thermo Fisher e coli bl21
    KHSRP is modified by SUMO1 at the major site K87 in vitro and in cells. a-b Exogenous and endogenous KHSRP in cells are modified by SUMO1. 293T cells transfected with indicated plasmids were lysed and pulled down with Ni 2+ -NTA resin for SUMOylation assay, and SUMO1 modification of KHSRP was analyzed by Western blotting with indicated antibodies. c SUMO1 modification of KHSRP is verified by in vitro E.coli -based SUMOylation assay. Plasmid pGEX-4T-1-KHSRP was co-transformed with or without pE1E2SUMO1 plasmid into E.coli <t>BL21</t> (DE3). After GST pull-down purification, Western blotting was conducted with anti-SUMO1 antibody and the same membrane was detected with anti-GST antibody after stripping. d Mutation of K87R weakens SUMO1 modification of KHSRP in 293T cells. The construct pEF-5HA-KHSRP-WT, or -K87R, or -K359R, or -K628R was co-transfected with His-SUMO1 into 293T cells. 48 h after transfection, cells were lysed for the SUMOylation assay with Ni 2+ -NTA resin
    E Coli Bl21, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli bl21/product/Thermo Fisher
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    e coli bl21 - by Bioz Stars, 2020-02
    99/100 stars
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    KHSRP is modified by SUMO1 at the major site K87 in vitro and in cells. a-b Exogenous and endogenous KHSRP in cells are modified by SUMO1. 293T cells transfected with indicated plasmids were lysed and pulled down with Ni 2+ -NTA resin for SUMOylation assay, and SUMO1 modification of KHSRP was analyzed by Western blotting with indicated antibodies. c SUMO1 modification of KHSRP is verified by in vitro E.coli -based SUMOylation assay. Plasmid pGEX-4T-1-KHSRP was co-transformed with or without pE1E2SUMO1 plasmid into E.coli BL21 (DE3). After GST pull-down purification, Western blotting was conducted with anti-SUMO1 antibody and the same membrane was detected with anti-GST antibody after stripping. d Mutation of K87R weakens SUMO1 modification of KHSRP in 293T cells. The construct pEF-5HA-KHSRP-WT, or -K87R, or -K359R, or -K628R was co-transfected with His-SUMO1 into 293T cells. 48 h after transfection, cells were lysed for the SUMOylation assay with Ni 2+ -NTA resin

    Journal: Molecular Cancer

    Article Title: SUMO1 modification of KHSRP regulates tumorigenesis by preventing the TL-G-Rich miRNA biogenesis

    doi: 10.1186/s12943-017-0724-6

    Figure Lengend Snippet: KHSRP is modified by SUMO1 at the major site K87 in vitro and in cells. a-b Exogenous and endogenous KHSRP in cells are modified by SUMO1. 293T cells transfected with indicated plasmids were lysed and pulled down with Ni 2+ -NTA resin for SUMOylation assay, and SUMO1 modification of KHSRP was analyzed by Western blotting with indicated antibodies. c SUMO1 modification of KHSRP is verified by in vitro E.coli -based SUMOylation assay. Plasmid pGEX-4T-1-KHSRP was co-transformed with or without pE1E2SUMO1 plasmid into E.coli BL21 (DE3). After GST pull-down purification, Western blotting was conducted with anti-SUMO1 antibody and the same membrane was detected with anti-GST antibody after stripping. d Mutation of K87R weakens SUMO1 modification of KHSRP in 293T cells. The construct pEF-5HA-KHSRP-WT, or -K87R, or -K359R, or -K628R was co-transfected with His-SUMO1 into 293T cells. 48 h after transfection, cells were lysed for the SUMOylation assay with Ni 2+ -NTA resin

    Article Snippet: Briefly, pGEX-4T-1-KHSRP-WT was co-expressed with or without pE1E2SUMO1 plasmid in E.coli BL21 (DE3) respectively, and then lysed by using B-PER Protein Extraction Reagent (#78248, Thermo Fisher, USA) and incubated wi th Glutathione sepharose 4B (GE healthcare) at 4 °C overnight.

    Techniques: Modification, In Vitro, Transfection, Western Blot, Plasmid Preparation, Transformation Assay, Purification, Stripping Membranes, Mutagenesis, Construct