bl21 cells  (Thermo Fisher)


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    Name:
    MultiShot StripWell BL21 Star DE3 Competent Cells
    Description:
    MultiShot StripWell BL21 Star DE3 chemically competent E coli cells are cloning competent cells that are packaged in a rack containing 12 strips of 8 wells to increase productivity for medium throughput bacterial transformations MultiShot StripWell BL21 Star DE3 competent E coli cells are designed for applications that require high level expression of non toxic recombinant proteins from low copy number T7 promoter based expression systems e g Champion pET vectors MultiShot StripWell BL21 Star DE3 Competent Cells are provided at a transformation efficiency of 1 x 107 cfu µg plasmid DNA Enhanced expression of nontoxic recombinant proteins MultiShot StripWell BL21 Star DE3 Competent Cells contain the DE3 lysogen that carries the gene for T7 RNA polymerase under control of the lacUV5 promoter IPTG is required to induce expression This strain also offers enhanced mRNA stability due to a mutation in the RNaseE gene rne131 that reduces levels of endogenous RNases and mRNA degradation thereby increasing the stability of mRNA transcripts and increasing protein yield Protein expression is further enhanced by the absence of the lon and outer membrane OmpT proteases which reduces degradation of heterologous proteins Key features of the BL21 Star DE3 Competent Cells include • rne131 for promotion of high mRNA stability and protein yield • OmpT and lack of lon for minimal degradation of heterologous proteins by proteases • Optimized for use with low copy number T7 promoter based plasmids It should be noted that BL21 Star strains have higher basal expression of heterologous genes than BL21 strains due to the increased stability of mRNA Therefore these strains may not be useful for expression of toxic genes MultiShot StripWell format flexible for no waste Each StripWell of eight connected tubes is designed with the same single tube single use features as our One Shot format tubes This format allows for all steps of the transformation protocol up to plating to take place in the same tube thereby saving time and to preventing contamination After addition of DNA MultiShot StripWell chemically competent cells can be transformed by heat shock at 42°C using a water bath all in the same tube The StripWell tubes can be cut to permit transformations in a single tube eliminating wasted aliquots of cells and avoiding freeze thaws that can result in reduced transformation efficiency Key features of the MultiShot StripWell format include • Increase productivity for medium throughput experimentation • Maximal flexibility transform as few as 1 tube and as many as 96 from the StripWell rack • Familiar convenience no aliquotting add DNA directly to cells heat shock and recover in the same tube • Peace of mind single use transformation minimizes contamination Genotype F ompT hsdSB rB mB gal dcm rne131 DE3 Find the strain and format that you need We offer many other E coli strains and formats of chemically competent cells and electrocompetent cells to help meet your specific transformation needs If you require other high throughput transformation options choose from our collection of MultiShot formatted comp cells or email us for custom formatting options
    Catalog Number:
    c609601
    Price:
    None
    Applications:
    Bacterial Expression|Cloning|Protein Biology|Protein Expression|Transformation
    Category:
    Competent Cells Strains
    Buy from Supplier


    Structured Review

    Thermo Fisher bl21 cells
    Characterization of MITF interactions with BAF60 subunits (A) HEK 293T cells were transfected with empty vector or a vector expressing 3X-Flag tagged BAF60A , BAF60B , or BAF60C . Cells were harvested after 48 hours. Whole cell extract was used for immunoprecipitation with IgG as a control or with FLAG antibody tagged Sigma M2 beads. The immunoprecipitated material was run on an SDS – polyacrylamide gel and immunoblotted for FLAG, BRG1, MITF, BAF60A (A301–595), BAF60B (A301–596, BAF60C (ab50566). (B) 293T cells were co-transfected with 3X-FLAG BAF60A and Mitf deletion constructs. Immunoprecipitations were performed with an antibody to FLAG and run on an SDS-polyacrylamide gel. Western blotting was performed with FLAG and MITF antibodies. Top: Schematic showing C terminal deletion constructs relative to the basic helix loop helix-leucine zipper (bHLH-LZ) and the C terminal transactivation domain from 324 to 369 shown in black. Middle: FLAG and MITF expression in cell extract (inputs). Bottom: FLAG and MITF expression detected from FLAG immunoprecipitations. (C) <t>BL21</t> bacterial cells were transformed with either an MITF plasmid or a plasmid expressing GST or GST-BAF60A, cultured, and induced with IPTG for 4 hours. GST or GST-BAF60A was immunoprecipitated with GST-agarose. The beads were then incubated with extract from MITF transformed cells and pulldowns were performed. Inputs are from MITF transformed cells.
    MultiShot StripWell BL21 Star DE3 chemically competent E coli cells are cloning competent cells that are packaged in a rack containing 12 strips of 8 wells to increase productivity for medium throughput bacterial transformations MultiShot StripWell BL21 Star DE3 competent E coli cells are designed for applications that require high level expression of non toxic recombinant proteins from low copy number T7 promoter based expression systems e g Champion pET vectors MultiShot StripWell BL21 Star DE3 Competent Cells are provided at a transformation efficiency of 1 x 107 cfu µg plasmid DNA Enhanced expression of nontoxic recombinant proteins MultiShot StripWell BL21 Star DE3 Competent Cells contain the DE3 lysogen that carries the gene for T7 RNA polymerase under control of the lacUV5 promoter IPTG is required to induce expression This strain also offers enhanced mRNA stability due to a mutation in the RNaseE gene rne131 that reduces levels of endogenous RNases and mRNA degradation thereby increasing the stability of mRNA transcripts and increasing protein yield Protein expression is further enhanced by the absence of the lon and outer membrane OmpT proteases which reduces degradation of heterologous proteins Key features of the BL21 Star DE3 Competent Cells include • rne131 for promotion of high mRNA stability and protein yield • OmpT and lack of lon for minimal degradation of heterologous proteins by proteases • Optimized for use with low copy number T7 promoter based plasmids It should be noted that BL21 Star strains have higher basal expression of heterologous genes than BL21 strains due to the increased stability of mRNA Therefore these strains may not be useful for expression of toxic genes MultiShot StripWell format flexible for no waste Each StripWell of eight connected tubes is designed with the same single tube single use features as our One Shot format tubes This format allows for all steps of the transformation protocol up to plating to take place in the same tube thereby saving time and to preventing contamination After addition of DNA MultiShot StripWell chemically competent cells can be transformed by heat shock at 42°C using a water bath all in the same tube The StripWell tubes can be cut to permit transformations in a single tube eliminating wasted aliquots of cells and avoiding freeze thaws that can result in reduced transformation efficiency Key features of the MultiShot StripWell format include • Increase productivity for medium throughput experimentation • Maximal flexibility transform as few as 1 tube and as many as 96 from the StripWell rack • Familiar convenience no aliquotting add DNA directly to cells heat shock and recover in the same tube • Peace of mind single use transformation minimizes contamination Genotype F ompT hsdSB rB mB gal dcm rne131 DE3 Find the strain and format that you need We offer many other E coli strains and formats of chemically competent cells and electrocompetent cells to help meet your specific transformation needs If you require other high throughput transformation options choose from our collection of MultiShot formatted comp cells or email us for custom formatting options
    https://www.bioz.com/result/bl21 cells/product/Thermo Fisher
    Average 99 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    bl21 cells - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "BAF60A mediates interactions between MITF and the BRG1-containing SWI/SNF complex during melanocyte differentiation"

    Article Title: BAF60A mediates interactions between MITF and the BRG1-containing SWI/SNF complex during melanocyte differentiation

    Journal: Journal of cellular physiology

    doi: 10.1002/jcp.27840

    Characterization of MITF interactions with BAF60 subunits (A) HEK 293T cells were transfected with empty vector or a vector expressing 3X-Flag tagged BAF60A , BAF60B , or BAF60C . Cells were harvested after 48 hours. Whole cell extract was used for immunoprecipitation with IgG as a control or with FLAG antibody tagged Sigma M2 beads. The immunoprecipitated material was run on an SDS – polyacrylamide gel and immunoblotted for FLAG, BRG1, MITF, BAF60A (A301–595), BAF60B (A301–596, BAF60C (ab50566). (B) 293T cells were co-transfected with 3X-FLAG BAF60A and Mitf deletion constructs. Immunoprecipitations were performed with an antibody to FLAG and run on an SDS-polyacrylamide gel. Western blotting was performed with FLAG and MITF antibodies. Top: Schematic showing C terminal deletion constructs relative to the basic helix loop helix-leucine zipper (bHLH-LZ) and the C terminal transactivation domain from 324 to 369 shown in black. Middle: FLAG and MITF expression in cell extract (inputs). Bottom: FLAG and MITF expression detected from FLAG immunoprecipitations. (C) BL21 bacterial cells were transformed with either an MITF plasmid or a plasmid expressing GST or GST-BAF60A, cultured, and induced with IPTG for 4 hours. GST or GST-BAF60A was immunoprecipitated with GST-agarose. The beads were then incubated with extract from MITF transformed cells and pulldowns were performed. Inputs are from MITF transformed cells.
    Figure Legend Snippet: Characterization of MITF interactions with BAF60 subunits (A) HEK 293T cells were transfected with empty vector or a vector expressing 3X-Flag tagged BAF60A , BAF60B , or BAF60C . Cells were harvested after 48 hours. Whole cell extract was used for immunoprecipitation with IgG as a control or with FLAG antibody tagged Sigma M2 beads. The immunoprecipitated material was run on an SDS – polyacrylamide gel and immunoblotted for FLAG, BRG1, MITF, BAF60A (A301–595), BAF60B (A301–596, BAF60C (ab50566). (B) 293T cells were co-transfected with 3X-FLAG BAF60A and Mitf deletion constructs. Immunoprecipitations were performed with an antibody to FLAG and run on an SDS-polyacrylamide gel. Western blotting was performed with FLAG and MITF antibodies. Top: Schematic showing C terminal deletion constructs relative to the basic helix loop helix-leucine zipper (bHLH-LZ) and the C terminal transactivation domain from 324 to 369 shown in black. Middle: FLAG and MITF expression in cell extract (inputs). Bottom: FLAG and MITF expression detected from FLAG immunoprecipitations. (C) BL21 bacterial cells were transformed with either an MITF plasmid or a plasmid expressing GST or GST-BAF60A, cultured, and induced with IPTG for 4 hours. GST or GST-BAF60A was immunoprecipitated with GST-agarose. The beads were then incubated with extract from MITF transformed cells and pulldowns were performed. Inputs are from MITF transformed cells.

    Techniques Used: Transfection, Plasmid Preparation, Expressing, Immunoprecipitation, Construct, Western Blot, Transformation Assay, Cell Culture, Incubation

    2) Product Images from "The DNA mimic protein BCAS0292 is involved in the regulation of virulence of Burkholderia cenocepacia"

    Article Title: The DNA mimic protein BCAS0292 is involved in the regulation of virulence of Burkholderia cenocepacia

    Journal: bioRxiv

    doi: 10.1101/2020.08.06.226779

    Role of BCAS0292 in epithelial cell attachment. A) Attachment of K56-2 WT, ΔBCAS0292 mutant and Δ0292_0292 complemented strain to CFBE41o − cells at an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain with 20 fields of view counted per data point in three independent experiments, ** p =0.0073. B) Confocal microscopy images representing the attachment of E. coli BL21 control and E. coli BL21_BCAS0292 strains to CFBE41o-cells. Bacteria were labelled using a primary anti- E. coli FITC-conjugated antibody (green). Nuclei of CFBE41o-cells were counterstained with DAPI (blue) and actin stained with phalloidin conjugated with Alexa fluor 568. C) Attachment E. coli BL21 control and E. coli BL21_BCAS0292 to CFBE41o-cells, using an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain, with 20 fields of view counted per data point in three independent experiments. D) Confirmation of expression of BCAS0292, following induction in E. coli BL21 cells with 1mM IPTG by Coomassie Blue stained SDS PAGE gel (conditions used for Figure 1C ). Lane 1: MW marker; Lane 2: E. coli BL21 expression control; Lane 3: E. coli BL21_BCAS0292 after incubation with IPTG (1mM) for 2 h.
    Figure Legend Snippet: Role of BCAS0292 in epithelial cell attachment. A) Attachment of K56-2 WT, ΔBCAS0292 mutant and Δ0292_0292 complemented strain to CFBE41o − cells at an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain with 20 fields of view counted per data point in three independent experiments, ** p =0.0073. B) Confocal microscopy images representing the attachment of E. coli BL21 control and E. coli BL21_BCAS0292 strains to CFBE41o-cells. Bacteria were labelled using a primary anti- E. coli FITC-conjugated antibody (green). Nuclei of CFBE41o-cells were counterstained with DAPI (blue) and actin stained with phalloidin conjugated with Alexa fluor 568. C) Attachment E. coli BL21 control and E. coli BL21_BCAS0292 to CFBE41o-cells, using an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain, with 20 fields of view counted per data point in three independent experiments. D) Confirmation of expression of BCAS0292, following induction in E. coli BL21 cells with 1mM IPTG by Coomassie Blue stained SDS PAGE gel (conditions used for Figure 1C ). Lane 1: MW marker; Lane 2: E. coli BL21 expression control; Lane 3: E. coli BL21_BCAS0292 after incubation with IPTG (1mM) for 2 h.

    Techniques Used: Cell Attachment Assay, Mutagenesis, Confocal Microscopy, Staining, Expressing, SDS Page, Marker, Incubation

    3) Product Images from "Rab18 binds to classical swine fever virus NS5A and mediates viral replication and assembly in swine umbilical vein endothelial cells"

    Article Title: Rab18 binds to classical swine fever virus NS5A and mediates viral replication and assembly in swine umbilical vein endothelial cells

    Journal: Virulence

    doi: 10.1080/21505594.2020.1767356

    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli BL21 (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Figure Legend Snippet: Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli BL21 (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.

    Techniques Used: Co-Immunoprecipitation Assay, Transfection, Immunoprecipitation, Western Blot, GST Pulldown Assay, Purification, Incubation, Expressing, Negative Control, Staining

    Related Articles

    Recombinant:

    Article Title: The DNA mimic protein BCAS0292 is involved in the regulation of virulence of Burkholderia cenocepacia
    Article Snippet: .. Following the confirmation of recombinant BCAS0292 (rBCAS0292) expression in a pilot study, 1 L cultures of BL21 cells transformed with the expression plasmid, were grown in LB-ampicillin (100 μg/ ml) and induced with IPTG at a final concentration of 1 mM overnight at 25 °C. .. Recombinant BCAS0292 purification Purification of rBCAS0292 was carried out as previously described ( ).

    Construct:

    Article Title: A dynamic regulatory interface on SARS-CoV-2 RNA polymerase
    Article Snippet: .. All constructs were transformed into Escherichia coli BL21(DE3) competent cells. ..

    Article Title: Versatile, Multivalent Nanobody Cocktails Efficiently Neutralize SARS-CoV-2
    Article Snippet: .. Expression and purification of proteinsNb DNA constructs were transformed into BL21(DE3) cells and plated on Agar with 50 μg/ml ampicillin at 37 °C overnight. .. Cells were cultured in an LB broth to reach an O.D. of ~ 0.5-0.6 before IPTG (0.5 mM) induct ion at 16°C overnight.

    Purification:

    Article Title: Versatile, Multivalent Nanobody Cocktails Efficiently Neutralize SARS-CoV-2
    Article Snippet: .. Expression and purification of proteinsNb DNA constructs were transformed into BL21(DE3) cells and plated on Agar with 50 μg/ml ampicillin at 37 °C overnight. .. Cells were cultured in an LB broth to reach an O.D. of ~ 0.5-0.6 before IPTG (0.5 mM) induct ion at 16°C overnight.

    Article Title: Golgi-resident TRIO regulates membrane trafficking during neurite outgrowth
    Article Snippet: .. Purification of the His6 -RABIN8 and direct binding assay His6 -RABIN8 were expressed in BL21(DE3) cells that had been induced with 1 m m IPTG at 37 °C for 4 h. Bacteria pellets were then sonicated in PBS with 1 mg/ml lysozyme. .. Proteins were then pelleted by centrifugation at 9000 rpm for 10 min and dissolved in PBS with 8 m urea.

    Concentration Assay:

    Article Title: The DNA mimic protein BCAS0292 is involved in the regulation of virulence of Burkholderia cenocepacia
    Article Snippet: .. Following the confirmation of recombinant BCAS0292 (rBCAS0292) expression in a pilot study, 1 L cultures of BL21 cells transformed with the expression plasmid, were grown in LB-ampicillin (100 μg/ ml) and induced with IPTG at a final concentration of 1 mM overnight at 25 °C. .. Recombinant BCAS0292 purification Purification of rBCAS0292 was carried out as previously described ( ).

    other:

    Article Title: Mapping RNAPII CTD Phosphorylation Reveals That the Identity and Modification of Seventh Heptad Residues Direct Tyr1 Phosphorylation
    Article Snippet: BL21 (DE3) cells coexpressing the human ABL1 kinase domain (residues 229−511) and YopH phosphatase from Yersinia were a kind gift from the Kuriyan lab.

    Expressing:

    Article Title: CRISPR/Cas9-based genome editing in the silverleaf whitefly (Bemisia tabaci)
    Article Snippet: .. Plasmids were transformed into BL21(DE3) cells for BtKV-Cas9/mCh expression following standard protein expression protocols. ..

    Article Title: The DNA mimic protein BCAS0292 is involved in the regulation of virulence of Burkholderia cenocepacia
    Article Snippet: .. Following the confirmation of recombinant BCAS0292 (rBCAS0292) expression in a pilot study, 1 L cultures of BL21 cells transformed with the expression plasmid, were grown in LB-ampicillin (100 μg/ ml) and induced with IPTG at a final concentration of 1 mM overnight at 25 °C. .. Recombinant BCAS0292 purification Purification of rBCAS0292 was carried out as previously described ( ).

    Article Title: Versatile, Multivalent Nanobody Cocktails Efficiently Neutralize SARS-CoV-2
    Article Snippet: .. Expression and purification of proteinsNb DNA constructs were transformed into BL21(DE3) cells and plated on Agar with 50 μg/ml ampicillin at 37 °C overnight. .. Cells were cultured in an LB broth to reach an O.D. of ~ 0.5-0.6 before IPTG (0.5 mM) induct ion at 16°C overnight.

    Sequencing:

    Article Title: Improving prognosis of surrogate assay for breast cancer patients by absolute quantitation of Ki67 protein levels using Quantitative Dot Blot (QDB) method
    Article Snippet: .. The plasmid was verified by sequencing,95 and expressed in BL21 (DE3) competent cells. .. The cells were induced with IPTG, and 96 total bacterial lysate was extracted in 10ml binding buffer (20 mM sodium phosphate, 97 500 mM NaCI, 20 mM imidazole, PH 7.4) before it was loaded onto a high affinity Ni2+ 98 column pre-equilibrated with 10ml binding buffer.

    Sonication:

    Article Title: Golgi-resident TRIO regulates membrane trafficking during neurite outgrowth
    Article Snippet: .. Purification of the His6 -RABIN8 and direct binding assay His6 -RABIN8 were expressed in BL21(DE3) cells that had been induced with 1 m m IPTG at 37 °C for 4 h. Bacteria pellets were then sonicated in PBS with 1 mg/ml lysozyme. .. Proteins were then pelleted by centrifugation at 9000 rpm for 10 min and dissolved in PBS with 8 m urea.

    Transformation Assay:

    Article Title: A dynamic regulatory interface on SARS-CoV-2 RNA polymerase
    Article Snippet: .. All constructs were transformed into Escherichia coli BL21(DE3) competent cells. ..

    Article Title: CRISPR/Cas9-based genome editing in the silverleaf whitefly (Bemisia tabaci)
    Article Snippet: .. Plasmids were transformed into BL21(DE3) cells for BtKV-Cas9/mCh expression following standard protein expression protocols. ..

    Article Title: The DNA mimic protein BCAS0292 is involved in the regulation of virulence of Burkholderia cenocepacia
    Article Snippet: .. Following the confirmation of recombinant BCAS0292 (rBCAS0292) expression in a pilot study, 1 L cultures of BL21 cells transformed with the expression plasmid, were grown in LB-ampicillin (100 μg/ ml) and induced with IPTG at a final concentration of 1 mM overnight at 25 °C. .. Recombinant BCAS0292 purification Purification of rBCAS0292 was carried out as previously described ( ).

    Article Title: Versatile, Multivalent Nanobody Cocktails Efficiently Neutralize SARS-CoV-2
    Article Snippet: .. Expression and purification of proteinsNb DNA constructs were transformed into BL21(DE3) cells and plated on Agar with 50 μg/ml ampicillin at 37 °C overnight. .. Cells were cultured in an LB broth to reach an O.D. of ~ 0.5-0.6 before IPTG (0.5 mM) induct ion at 16°C overnight.

    Article Title: Glucosamine-6P and glucosamine-1P, respectively an activator and a substrate of rhodococcal ADP-glucose pyrophosphorylases, show a hint to ascertain (actino)bacterial glucosamine metabolism
    Article Snippet: .. Competent E. coli BL21 (DE3) cells were transformed with the [pET28c/RfaglgC ] construction. .. Protein expression was carried out using LB medium (10 g/l tryptone; 5 g/l yeast extract; 5 g/l NaCl) supplemented with 100 g/ml kanamycin.

    Binding Assay:

    Article Title: Golgi-resident TRIO regulates membrane trafficking during neurite outgrowth
    Article Snippet: .. Purification of the His6 -RABIN8 and direct binding assay His6 -RABIN8 were expressed in BL21(DE3) cells that had been induced with 1 m m IPTG at 37 °C for 4 h. Bacteria pellets were then sonicated in PBS with 1 mg/ml lysozyme. .. Proteins were then pelleted by centrifugation at 9000 rpm for 10 min and dissolved in PBS with 8 m urea.

    Plasmid Preparation:

    Article Title: Improving prognosis of surrogate assay for breast cancer patients by absolute quantitation of Ki67 protein levels using Quantitative Dot Blot (QDB) method
    Article Snippet: .. The plasmid was verified by sequencing,95 and expressed in BL21 (DE3) competent cells. .. The cells were induced with IPTG, and 96 total bacterial lysate was extracted in 10ml binding buffer (20 mM sodium phosphate, 97 500 mM NaCI, 20 mM imidazole, PH 7.4) before it was loaded onto a high affinity Ni2+ 98 column pre-equilibrated with 10ml binding buffer.

    Article Title: The DNA mimic protein BCAS0292 is involved in the regulation of virulence of Burkholderia cenocepacia
    Article Snippet: .. Following the confirmation of recombinant BCAS0292 (rBCAS0292) expression in a pilot study, 1 L cultures of BL21 cells transformed with the expression plasmid, were grown in LB-ampicillin (100 μg/ ml) and induced with IPTG at a final concentration of 1 mM overnight at 25 °C. .. Recombinant BCAS0292 purification Purification of rBCAS0292 was carried out as previously described ( ).

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  • 99
    Thermo Fisher e coli bl21
    Spot assays used for monitoring the growth performance of <t>BL21/pEZYHb</t> and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress
    E Coli Bl21, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 151 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 151 article reviews
    Price from $9.99 to $1999.99
    e coli bl21 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Thermo Fisher e coli bl21 cells
    SDS-PAGE analysis and activity of nitrite reductase expressed by E. coli <t>BL21.</t> (a) M: protein marker; Lane 1: protein product. (b) Nitrite reductase activity of E. coli BL21 transformed via OMVs under different temperatures.
    E Coli Bl21 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli bl21 cells/product/Thermo Fisher
    Average 92 stars, based on 39 article reviews
    Price from $9.99 to $1999.99
    e coli bl21 cells - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    Image Search Results


    Spot assays used for monitoring the growth performance of BL21/pEZYHb and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress

    Journal: BMC Genomics

    Article Title: The alcohol dehydrogenase gene family in sugarcane and its involvement in cold stress regulation

    doi: 10.1186/s12864-020-06929-9

    Figure Lengend Snippet: Spot assays used for monitoring the growth performance of BL21/pEZYHb and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress

    Article Snippet: E. coli BL21 with the empty vector pEZYHb was used as the control.

    Techniques: Transformation Assay, Incubation

    SDS-PAGE analysis and activity of nitrite reductase expressed by E. coli BL21. (a) M: protein marker; Lane 1: protein product. (b) Nitrite reductase activity of E. coli BL21 transformed via OMVs under different temperatures.

    Journal: bioRxiv

    Article Title: Outer Membrane Vesicles Mediated Horizontal Transfer of an Aerobic Denitrification Gene between Escherichia coli

    doi: 10.1101/835694

    Figure Lengend Snippet: SDS-PAGE analysis and activity of nitrite reductase expressed by E. coli BL21. (a) M: protein marker; Lane 1: protein product. (b) Nitrite reductase activity of E. coli BL21 transformed via OMVs under different temperatures.

    Article Snippet: E. coli BL21 cells were stained with VybrantTM DiO live cell tracer (ThermoFisher).

    Techniques: SDS Page, Activity Assay, Marker, Transformation Assay

    The LCSM images of E. coli BL21 induced by IPTG. PH: phase contrast image; Green fluorescence: green fluorescence image; Overlay: image of phase contrast image and green fluorescence image superimposed.

    Journal: bioRxiv

    Article Title: Outer Membrane Vesicles Mediated Horizontal Transfer of an Aerobic Denitrification Gene between Escherichia coli

    doi: 10.1101/835694

    Figure Lengend Snippet: The LCSM images of E. coli BL21 induced by IPTG. PH: phase contrast image; Green fluorescence: green fluorescence image; Overlay: image of phase contrast image and green fluorescence image superimposed.

    Article Snippet: E. coli BL21 cells were stained with VybrantTM DiO live cell tracer (ThermoFisher).

    Techniques: Confocal Laser Scanning Microscopy, Fluorescence

    Characterization of MITF interactions with BAF60 subunits (A) HEK 293T cells were transfected with empty vector or a vector expressing 3X-Flag tagged BAF60A , BAF60B , or BAF60C . Cells were harvested after 48 hours. Whole cell extract was used for immunoprecipitation with IgG as a control or with FLAG antibody tagged Sigma M2 beads. The immunoprecipitated material was run on an SDS – polyacrylamide gel and immunoblotted for FLAG, BRG1, MITF, BAF60A (A301–595), BAF60B (A301–596, BAF60C (ab50566). (B) 293T cells were co-transfected with 3X-FLAG BAF60A and Mitf deletion constructs. Immunoprecipitations were performed with an antibody to FLAG and run on an SDS-polyacrylamide gel. Western blotting was performed with FLAG and MITF antibodies. Top: Schematic showing C terminal deletion constructs relative to the basic helix loop helix-leucine zipper (bHLH-LZ) and the C terminal transactivation domain from 324 to 369 shown in black. Middle: FLAG and MITF expression in cell extract (inputs). Bottom: FLAG and MITF expression detected from FLAG immunoprecipitations. (C) BL21 bacterial cells were transformed with either an MITF plasmid or a plasmid expressing GST or GST-BAF60A, cultured, and induced with IPTG for 4 hours. GST or GST-BAF60A was immunoprecipitated with GST-agarose. The beads were then incubated with extract from MITF transformed cells and pulldowns were performed. Inputs are from MITF transformed cells.

    Journal: Journal of cellular physiology

    Article Title: BAF60A mediates interactions between MITF and the BRG1-containing SWI/SNF complex during melanocyte differentiation

    doi: 10.1002/jcp.27840

    Figure Lengend Snippet: Characterization of MITF interactions with BAF60 subunits (A) HEK 293T cells were transfected with empty vector or a vector expressing 3X-Flag tagged BAF60A , BAF60B , or BAF60C . Cells were harvested after 48 hours. Whole cell extract was used for immunoprecipitation with IgG as a control or with FLAG antibody tagged Sigma M2 beads. The immunoprecipitated material was run on an SDS – polyacrylamide gel and immunoblotted for FLAG, BRG1, MITF, BAF60A (A301–595), BAF60B (A301–596, BAF60C (ab50566). (B) 293T cells were co-transfected with 3X-FLAG BAF60A and Mitf deletion constructs. Immunoprecipitations were performed with an antibody to FLAG and run on an SDS-polyacrylamide gel. Western blotting was performed with FLAG and MITF antibodies. Top: Schematic showing C terminal deletion constructs relative to the basic helix loop helix-leucine zipper (bHLH-LZ) and the C terminal transactivation domain from 324 to 369 shown in black. Middle: FLAG and MITF expression in cell extract (inputs). Bottom: FLAG and MITF expression detected from FLAG immunoprecipitations. (C) BL21 bacterial cells were transformed with either an MITF plasmid or a plasmid expressing GST or GST-BAF60A, cultured, and induced with IPTG for 4 hours. GST or GST-BAF60A was immunoprecipitated with GST-agarose. The beads were then incubated with extract from MITF transformed cells and pulldowns were performed. Inputs are from MITF transformed cells.

    Article Snippet: For GST pulldowns, BL21 cells (Thermofisher, Waltham, MA, USA) were transformed with empty pGEX, pGEX-2tBAF60A, or pRSET-B-MITF.

    Techniques: Transfection, Plasmid Preparation, Expressing, Immunoprecipitation, Construct, Western Blot, Transformation Assay, Cell Culture, Incubation

    Detection of recombinant plasmids and expression and purification of recombinant proteins M1: DNA marker (DL2000); lane 1: PCR products of candidate antigen genes; lane 2: enzyme digestion of recombinant plasmids with Bam HI + Xho I; lane 3: enzyme digestion of recombinant plasmids with Bam HI; M2: DNA marker (DL1000); M3: DNA marker (DL10000); M4: DNA marker (DL15000). M: Protein markers. A1~A9: uninduced BL21 (pET32a) supernatant, uninduced BL21 (pET32a) sediment, induced BL21 (pET32a) supernatant, induced BL21 (pET32a) sediment, uninduced BL21 (P-Srr) supernatant, uninduced BL21 (P-Srr) sediment, induced BL21 (P-Srr) supernatant, induced BL21 (P-Srr) sediment, purification of recombinant rSrr. B1~B7: uninduced BL21 (pET32a), induced BL21 (pET32a), uninduced BL21 (P-NeuA), induced BL21 (P-NeuA), induced BL21 (P-NeuA) supernatant, induced BL21 (P-NeuA) sediment, purification of recombinant rNeuA. C1~C7: uninduced BL21 (pET32a), induced BL21 (pET32a), uninduced BL21 (P-Hsp), induced BL21 (P-Hsp), induced BL21 (P-Hsp) supernatant, induced BL21 (P-Hsp) sediment, purification of recombinant rHsp.

    Journal: Oncotarget

    Article Title: Identification and screening of effective protective antigens for channel catfish against Streptococcus iniae

    doi: 10.18632/oncotarget.16475

    Figure Lengend Snippet: Detection of recombinant plasmids and expression and purification of recombinant proteins M1: DNA marker (DL2000); lane 1: PCR products of candidate antigen genes; lane 2: enzyme digestion of recombinant plasmids with Bam HI + Xho I; lane 3: enzyme digestion of recombinant plasmids with Bam HI; M2: DNA marker (DL1000); M3: DNA marker (DL10000); M4: DNA marker (DL15000). M: Protein markers. A1~A9: uninduced BL21 (pET32a) supernatant, uninduced BL21 (pET32a) sediment, induced BL21 (pET32a) supernatant, induced BL21 (pET32a) sediment, uninduced BL21 (P-Srr) supernatant, uninduced BL21 (P-Srr) sediment, induced BL21 (P-Srr) supernatant, induced BL21 (P-Srr) sediment, purification of recombinant rSrr. B1~B7: uninduced BL21 (pET32a), induced BL21 (pET32a), uninduced BL21 (P-NeuA), induced BL21 (P-NeuA), induced BL21 (P-NeuA) supernatant, induced BL21 (P-NeuA) sediment, purification of recombinant rNeuA. C1~C7: uninduced BL21 (pET32a), induced BL21 (pET32a), uninduced BL21 (P-Hsp), induced BL21 (P-Hsp), induced BL21 (P-Hsp) supernatant, induced BL21 (P-Hsp) sediment, purification of recombinant rHsp.

    Article Snippet: The plasmids were then transformed into E. coli BL21 cells and induced by adding 1.0 mM IPTG at 37°C for 4 h. The cells were then centrifuged at 8000 × g for 10 min at 4°C, suspended in sterile phosphate-buffered saline (PBS), sonicated with a Sonic Dismembrator (model 500; Thermo Fisher Scientific, Waltham, MA, USA), and examined by 12.5% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

    Techniques: Recombinant, Expressing, Purification, Marker, Polymerase Chain Reaction