bl21  (New England Biolabs)


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    Name:
    BL21 DE3 Competent E coli
    Description:
    BL21 DE3 Competent E coli 20x0 05 ml
    Catalog Number:
    C2527H
    Price:
    204
    Category:
    Competent Bacteria
    Size:
    1 ml
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    Structured Review

    New England Biolabs bl21
    BL21 DE3 Competent E coli
    BL21 DE3 Competent E coli 20x0 05 ml
    https://www.bioz.com/result/bl21/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bl21 - by Bioz Stars, 2021-05
    99/100 stars

    Images

    1) Product Images from "The traffic ATPase PilF interacts with the inner membrane platform of the DNA translocator and type IV pili from Thermus thermophilus"

    Article Title: The traffic ATPase PilF interacts with the inner membrane platform of the DNA translocator and type IV pili from Thermus thermophilus

    Journal: FEBS Open Bio

    doi: 10.1002/2211-5463.12548

    Purification of PilMNO, PilMN, PilM, and PilN. His 6 ‐PilMNO‐Strep (lane 1), His 6 ‐PilMN‐Strep (lane 2), His 6 ‐PilM (lane 3), and PilN‐Strep (lane 4) were purified from Escherichia coli BL21 (DE3) cells as described in Experimental procedures . PilMNO, PilMN, and PilN were isolated from membranes, whereas PilM was isolated from soluble fractions. To separate His 6 ‐PilM (44 kDa), PilN (23 kDa), and PilO‐Strep (22.5 kDa), 4–20% SDS/PAGE was performed (lane 1). All other protein preparations were separated by 14% SDS/PAGE (A). The identity of the proteins was confirmed by western blot using polyclonal antisera against PilM (B) and PilN (C) or using Strep‐Tactin HRP conjugate to detect PilO‐Strep (D). Dotted lines indicate excised lanes from different but comparable gels or blots.
    Figure Legend Snippet: Purification of PilMNO, PilMN, PilM, and PilN. His 6 ‐PilMNO‐Strep (lane 1), His 6 ‐PilMN‐Strep (lane 2), His 6 ‐PilM (lane 3), and PilN‐Strep (lane 4) were purified from Escherichia coli BL21 (DE3) cells as described in Experimental procedures . PilMNO, PilMN, and PilN were isolated from membranes, whereas PilM was isolated from soluble fractions. To separate His 6 ‐PilM (44 kDa), PilN (23 kDa), and PilO‐Strep (22.5 kDa), 4–20% SDS/PAGE was performed (lane 1). All other protein preparations were separated by 14% SDS/PAGE (A). The identity of the proteins was confirmed by western blot using polyclonal antisera against PilM (B) and PilN (C) or using Strep‐Tactin HRP conjugate to detect PilO‐Strep (D). Dotted lines indicate excised lanes from different but comparable gels or blots.

    Techniques Used: Purification, Isolation, SDS Page, Western Blot

    Related Articles

    Transformation Assay:

    Article Title: Scale-up fermentation of Escherichia coli for the production of recombinant endoglucanase from Clostridium thermocellum
    Article Snippet: The ligation mixture was incubated on ice for 1 h and then at 20 °C for overnight. .. Competent cells of E. coli BL21 (DE3) were then transformed with this ligation mixture containing desired heterologous plasmid by conventional heat shock method. .. For the screening of transformants, the transformed cells were spread over the LB-agar plates (1.5% agar containing 50 μg ml−1 ampicillin) and was incubated at 37℃ overnight.

    Article Title: Identification of the agr Peptide of Listeria monocytogenes
    Article Snippet: 180 μl aliquots were distributed in 96 well microtiter plates (each condition in triplicate) and incubated at 30°C for 2 h. At this stage, 20 μl of diluted peptides were added to obtain the indicated final concentrations (5 nM–50 μM) and plates were incubated at 30°C in a Tecan Infinite M200 plate reader with hourly OD600 and luminescence intensity measurements. .. AIP Production in E. coli For heterologous AIP production, pET29a_agrB or pET29a_agrBD were transformed into E. coli BL21(DE3) (New England Biolabs) and transformants were selected on LB agar containing kanamycin. ..

    Ligation:

    Article Title: Scale-up fermentation of Escherichia coli for the production of recombinant endoglucanase from Clostridium thermocellum
    Article Snippet: The ligation mixture was incubated on ice for 1 h and then at 20 °C for overnight. .. Competent cells of E. coli BL21 (DE3) were then transformed with this ligation mixture containing desired heterologous plasmid by conventional heat shock method. .. For the screening of transformants, the transformed cells were spread over the LB-agar plates (1.5% agar containing 50 μg ml−1 ampicillin) and was incubated at 37℃ overnight.

    Plasmid Preparation:

    Article Title: Scale-up fermentation of Escherichia coli for the production of recombinant endoglucanase from Clostridium thermocellum
    Article Snippet: The ligation mixture was incubated on ice for 1 h and then at 20 °C for overnight. .. Competent cells of E. coli BL21 (DE3) were then transformed with this ligation mixture containing desired heterologous plasmid by conventional heat shock method. .. For the screening of transformants, the transformed cells were spread over the LB-agar plates (1.5% agar containing 50 μg ml−1 ampicillin) and was incubated at 37℃ overnight.

    Article Title: Cytoplasmic Chaperones Enhance Soluble Expression of Anti-EGFR huscFv in Escherichia coli
    Article Snippet: Objectives The aim of this study was to increase the expression of soluble humanized anti-EGFR scFv by co-expression with cytoplasmic chaperones under different conditions and to evaluate its activity on the EGFR-overexpressing tumor cell line A431. .. Cell lines, ScFv Construct, Chaperone Plasmids The E. coli strains, including BL21 (DE3), SHuffle® T7 Express Competent (NEB), BL21 Rosetta DE3, Inv, and the expression vector pET22b (+) were purchased from Novagen and New England Biolabs, respectively. .. The chaperone plasmid set was purchased from Takara Bio Inc. ( ). pET22b - humanized anti-EGFR ScFv construct was designed by Veisi K et al.( ) ( ).

    Article Title: A Multiprotein DNA Translocation Complex Directs Intramycelial Plasmid Spreading during Streptomyces Conjugation
    Article Snippet: .. The resulting expression plasmid pGMT7-spdB2 was used to transform the different E. coli strains [BL21(DE3), BL21(DE3)/pLysS, Lemo21(DE3) (NEB)]. .. As a control, cells were also transformed with the empty expression plasmid pGM202T7.

    Construct:

    Article Title: Cytoplasmic Chaperones Enhance Soluble Expression of Anti-EGFR huscFv in Escherichia coli
    Article Snippet: Objectives The aim of this study was to increase the expression of soluble humanized anti-EGFR scFv by co-expression with cytoplasmic chaperones under different conditions and to evaluate its activity on the EGFR-overexpressing tumor cell line A431. .. Cell lines, ScFv Construct, Chaperone Plasmids The E. coli strains, including BL21 (DE3), SHuffle® T7 Express Competent (NEB), BL21 Rosetta DE3, Inv, and the expression vector pET22b (+) were purchased from Novagen and New England Biolabs, respectively. .. The chaperone plasmid set was purchased from Takara Bio Inc. ( ). pET22b - humanized anti-EGFR ScFv construct was designed by Veisi K et al.( ) ( ).

    Expressing:

    Article Title: Cytoplasmic Chaperones Enhance Soluble Expression of Anti-EGFR huscFv in Escherichia coli
    Article Snippet: Objectives The aim of this study was to increase the expression of soluble humanized anti-EGFR scFv by co-expression with cytoplasmic chaperones under different conditions and to evaluate its activity on the EGFR-overexpressing tumor cell line A431. .. Cell lines, ScFv Construct, Chaperone Plasmids The E. coli strains, including BL21 (DE3), SHuffle® T7 Express Competent (NEB), BL21 Rosetta DE3, Inv, and the expression vector pET22b (+) were purchased from Novagen and New England Biolabs, respectively. .. The chaperone plasmid set was purchased from Takara Bio Inc. ( ). pET22b - humanized anti-EGFR ScFv construct was designed by Veisi K et al.( ) ( ).

    Article Title: A Multiprotein DNA Translocation Complex Directs Intramycelial Plasmid Spreading during Streptomyces Conjugation
    Article Snippet: .. The resulting expression plasmid pGMT7-spdB2 was used to transform the different E. coli strains [BL21(DE3), BL21(DE3)/pLysS, Lemo21(DE3) (NEB)]. .. As a control, cells were also transformed with the empty expression plasmid pGM202T7.

    Article Title: Molecular characterization and Functional Analysis of the PilQ380-706: a Novel Secretin Domain in Pseudomonas aeruginosa
    Article Snippet: .. Bacterial strains, plasmids, and media Escherichia coli (E. coli ) strains Top10F and BL21 (DE3) were used as preservation and expression hosts. .. The P. aeruginosa laboratory strain PAO1 (that kindly provided by Dr. Abdi from Department of Microbiology, Faculty of Biological Sciences, Alzahra University, Tehran, Iran) were performed.

    Preserving:

    Article Title: Molecular characterization and Functional Analysis of the PilQ380-706: a Novel Secretin Domain in Pseudomonas aeruginosa
    Article Snippet: .. Bacterial strains, plasmids, and media Escherichia coli (E. coli ) strains Top10F and BL21 (DE3) were used as preservation and expression hosts. .. The P. aeruginosa laboratory strain PAO1 (that kindly provided by Dr. Abdi from Department of Microbiology, Faculty of Biological Sciences, Alzahra University, Tehran, Iran) were performed.

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    New England Biolabs e coli bl21 strains
    Functional validation of the predicted ARGs and structural investigation of conserved sites detected by HMD-ARG. a Left figure: A diagram showing the procedure of heterologous expression and functional analysis of the predicted ARGs from PA150567 in E. coli <t>BL21</t> host. Middle figure : Growth curves of E. coli hosts with expression of the predicted β -lactamases that inactivate β -lactam antibiotics in the presence of 2 μg/ml meropenem. Right figure : Growth curves of E. coli hosts with expression of the predicted acetyltransferases that inactivate aminoglycoside antibiotics in the presence of 4 μg/ml amikacin. b The HMD-ARG predicted conserved sites and the corresponding sequence logo from 319 to 393 in AXX01_04100. c After we mutated the conserved site (346) from T to N, the mutated protein’s (colored in red) local structure (predicted by RaptorX) changed significantly from the wild type (colored in light blue). Moreover, the binding affinity (predicted by AutoDock) between the mutated protein and the ligand (antibiotics) also reduced, as illustrated in the middle (wild type) and right (mutated protein) figures
    E Coli Bl21 Strains, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli bl21 strains/product/New England Biolabs
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    Functional validation of the predicted ARGs and structural investigation of conserved sites detected by HMD-ARG. a Left figure: A diagram showing the procedure of heterologous expression and functional analysis of the predicted ARGs from PA150567 in E. coli BL21 host. Middle figure : Growth curves of E. coli hosts with expression of the predicted β -lactamases that inactivate β -lactam antibiotics in the presence of 2 μg/ml meropenem. Right figure : Growth curves of E. coli hosts with expression of the predicted acetyltransferases that inactivate aminoglycoside antibiotics in the presence of 4 μg/ml amikacin. b The HMD-ARG predicted conserved sites and the corresponding sequence logo from 319 to 393 in AXX01_04100. c After we mutated the conserved site (346) from T to N, the mutated protein’s (colored in red) local structure (predicted by RaptorX) changed significantly from the wild type (colored in light blue). Moreover, the binding affinity (predicted by AutoDock) between the mutated protein and the ligand (antibiotics) also reduced, as illustrated in the middle (wild type) and right (mutated protein) figures

    Journal: Microbiome

    Article Title: HMD-ARG: hierarchical multi-task deep learning for annotating antibiotic resistance genes

    doi: 10.1186/s40168-021-01002-3

    Figure Lengend Snippet: Functional validation of the predicted ARGs and structural investigation of conserved sites detected by HMD-ARG. a Left figure: A diagram showing the procedure of heterologous expression and functional analysis of the predicted ARGs from PA150567 in E. coli BL21 host. Middle figure : Growth curves of E. coli hosts with expression of the predicted β -lactamases that inactivate β -lactam antibiotics in the presence of 2 μg/ml meropenem. Right figure : Growth curves of E. coli hosts with expression of the predicted acetyltransferases that inactivate aminoglycoside antibiotics in the presence of 4 μg/ml amikacin. b The HMD-ARG predicted conserved sites and the corresponding sequence logo from 319 to 393 in AXX01_04100. c After we mutated the conserved site (346) from T to N, the mutated protein’s (colored in red) local structure (predicted by RaptorX) changed significantly from the wild type (colored in light blue). Moreover, the binding affinity (predicted by AutoDock) between the mutated protein and the ligand (antibiotics) also reduced, as illustrated in the middle (wild type) and right (mutated protein) figures

    Article Snippet: Overnight culture of E. coli BL21 strains containing the plasmids for overexpression of the predicted genes were 1:100 diluted and grown in LB medium supplemented with kanamycin (20 μg/ml) and Isopropyl β-D-1-thiogalactopyranoside (IPTG) (0.5 mM).

    Techniques: Functional Assay, Expressing, Sequencing, Binding Assay

    Use of THP-1 TLR4-CD14 NF-κB-eGFP reporter cells for the detection of microbial contaminations in recombinant protein preparations. (A) THP-1 TLR4-CD14 NF-κB-eGFP reporter cells were incubated with the indicated serial dilutions of ultrapure LPS for 24 h. NF-κB-driven eGFP expression was assessed by flow cytometry. Bar graphs show geometric mean of fluorescence intensity (gMFI). eGFP expression of the parental THP-1 reporter cells treated with 300 ng/ml ultrapure LPS is shown for comparison (grey bar). (B) mRNA expression of TLR4, CD14 and MD2 was measured in parental and TLR4-CD14 THP-1 reporter cells by real-time qPCR. Data are presented as mean ± SEM of expression values normalized against the housekeeping gene GAPDH (n = 4). (C) Recombinant human split product C4dg produced in three different expression systems (HEK293-6E, standard E . coli BL21 and E . coli ClearColi BL21) was tested for TLR4-agonist contaminations using HEK293 hTLR4A-MD2-CD14 cells. Following 24 h of incubation the IL-8 content in the culture supernatants was measured by ELISA. (D) Parental and TLR4-CD14 THP-1 reporters were incubated with the protein preparations described in (C) and NF-κB-driven eGFP expression was assessed by flow cytometry 24 h later. Bar graphs show geometric mean of fluorescence intensity (gMFI). Mean and SE were calculated from duplicates of four independently performed experiments (n = 4) (E) Immature human moDCs were incubated with standard LPS, E . coli BL21 or E . coli ClearColi BL21 expressed C4dg protein at the indicated concentrations for 24 h or were left untreated. Expression of maturation markers CD83 and CD86 was assessed by flow cytometry. (F) E . coli ClearColi BL21-expressed C4dg protein was subjected to a single round of bulk chromatography using a polymyxin resin (see Material and Methods ). Samples before and after chromatography were tested using the THP-1 TLR4-CD14 reporters. Mean and SE were calculated from duplicates of four independently performed experiments (n = 4).

    Journal: PLoS ONE

    Article Title: A human monocytic NF-κB fluorescent reporter cell line for detection of microbial contaminants in biological samples

    doi: 10.1371/journal.pone.0178220

    Figure Lengend Snippet: Use of THP-1 TLR4-CD14 NF-κB-eGFP reporter cells for the detection of microbial contaminations in recombinant protein preparations. (A) THP-1 TLR4-CD14 NF-κB-eGFP reporter cells were incubated with the indicated serial dilutions of ultrapure LPS for 24 h. NF-κB-driven eGFP expression was assessed by flow cytometry. Bar graphs show geometric mean of fluorescence intensity (gMFI). eGFP expression of the parental THP-1 reporter cells treated with 300 ng/ml ultrapure LPS is shown for comparison (grey bar). (B) mRNA expression of TLR4, CD14 and MD2 was measured in parental and TLR4-CD14 THP-1 reporter cells by real-time qPCR. Data are presented as mean ± SEM of expression values normalized against the housekeeping gene GAPDH (n = 4). (C) Recombinant human split product C4dg produced in three different expression systems (HEK293-6E, standard E . coli BL21 and E . coli ClearColi BL21) was tested for TLR4-agonist contaminations using HEK293 hTLR4A-MD2-CD14 cells. Following 24 h of incubation the IL-8 content in the culture supernatants was measured by ELISA. (D) Parental and TLR4-CD14 THP-1 reporters were incubated with the protein preparations described in (C) and NF-κB-driven eGFP expression was assessed by flow cytometry 24 h later. Bar graphs show geometric mean of fluorescence intensity (gMFI). Mean and SE were calculated from duplicates of four independently performed experiments (n = 4) (E) Immature human moDCs were incubated with standard LPS, E . coli BL21 or E . coli ClearColi BL21 expressed C4dg protein at the indicated concentrations for 24 h or were left untreated. Expression of maturation markers CD83 and CD86 was assessed by flow cytometry. (F) E . coli ClearColi BL21-expressed C4dg protein was subjected to a single round of bulk chromatography using a polymyxin resin (see Material and Methods ). Samples before and after chromatography were tested using the THP-1 TLR4-CD14 reporters. Mean and SE were calculated from duplicates of four independently performed experiments (n = 4).

    Article Snippet: The electrocompetent E. coli protein expression strain ClearColi BL21 was purchased from Lucigen (Middleton, WI) and standard E. coli BL21 were obtained from New England Biolabs (Ipswich, MA).

    Techniques: Recombinant, Incubation, Expressing, Flow Cytometry, Cytometry, Fluorescence, Real-time Polymerase Chain Reaction, Produced, Enzyme-linked Immunosorbent Assay, Chromatography

    Sender plasmids expressed in Escherichia coli BL21. (A) The modular Sender vector allows facile cloning of any EcoRI , XbaI ] glyphs (top) and a scaled circular map (bottom). (B) Optical density (OD) readings for E . coli cultures expressing each of the Sender plasmids. (C) mCherry expression is shown as end-point RFP signal normalized to OD 600 after an 8 hour growth period.

    Journal: PLoS ONE

    Article Title: Characterization of diverse homoserine lactone synthases in Escherichia coli

    doi: 10.1371/journal.pone.0202294

    Figure Lengend Snippet: Sender plasmids expressed in Escherichia coli BL21. (A) The modular Sender vector allows facile cloning of any EcoRI , XbaI ] glyphs (top) and a scaled circular map (bottom). (B) Optical density (OD) readings for E . coli cultures expressing each of the Sender plasmids. (C) mCherry expression is shown as end-point RFP signal normalized to OD 600 after an 8 hour growth period.

    Article Snippet: Cultures of E . coli BL21 (New England Biolabs) transformed with Sender plasmids, a Receiver plasmid, a negative receiver plasmid, and a GFP positive plasmid were grown in 3 mL of LB with 5 ug/mL ampicillin for 16 hours at 37°C and shaking at 220 rpm.

    Techniques: Plasmid Preparation, Clone Assay, Expressing

    ( a) The SDS-PAGE analysis of protein tal and dxs expressed in E. coli with and without tags designed by our optimization algorithm. “+” and “-” represented expressed proteins with and without peptide tags respectively. “P” and “S” represented the pellet fraction (insoluble) and supernatant fraction (soluble), respectively. The oval shapes highlight the bands of dxs and tal proteins. Protein tal and dxs were expressed in K3 medium with 20 g/L glucose at 30 °C. ( b) Quantitative presentation of the SDS-PAGE images in a . The protein solubility was the ratio of soluble protein amount to the total protein amount. The protein amount was estimated by using band intensity on SDS-PAGE images. The sequences of the designed tags for N-terminal and C-terminal were shown. The amino acid S and G on the two ends of the tags were the linkers for GT DNA assembly standard, which was used to construct the plasmids in this study 40 .

    Journal: bioRxiv

    Article Title: Improve Protein Solubility and Activity based on Machine Learning Models

    doi: 10.1101/817890

    Figure Lengend Snippet: ( a) The SDS-PAGE analysis of protein tal and dxs expressed in E. coli with and without tags designed by our optimization algorithm. “+” and “-” represented expressed proteins with and without peptide tags respectively. “P” and “S” represented the pellet fraction (insoluble) and supernatant fraction (soluble), respectively. The oval shapes highlight the bands of dxs and tal proteins. Protein tal and dxs were expressed in K3 medium with 20 g/L glucose at 30 °C. ( b) Quantitative presentation of the SDS-PAGE images in a . The protein solubility was the ratio of soluble protein amount to the total protein amount. The protein amount was estimated by using band intensity on SDS-PAGE images. The sequences of the designed tags for N-terminal and C-terminal were shown. The amino acid S and G on the two ends of the tags were the linkers for GT DNA assembly standard, which was used to construct the plasmids in this study 40 .

    Article Snippet: Cell culture and SDS-PAGE analysis of protein solubility Each of constructed plasmid was introduced into E. coli BL21 (DE3) (C2530H, New England Biolabs) for SDS-PAGE analysis by using standard heat shock protocol.

    Techniques: SDS Page, Solubility, Construct