Structured Review

Millipore bl21 de3
mFIZZ1 soluble expressed using wheat germ extract. ( A ) The expression of mFIZZ1 in SHuffle™ T7, Origami <t>DE3</t> and <t>BL21</t> DE3 analysed on non-reducing 15% SDS-PAGE stained with Coomassie Brilliant Blue. ( B ) Strip of the immunoblot from the SDS-PAGE in ( A ) developed with anti-His antibody is shown. ( C ) The expression of mFIZZ1′ with wheat germ extract analysed on non-reducing 15% SDS-PAGE stained with Coomassie Brilliant Blue. ( D ) Strip of the respective immunoblot of ( C ) developed with anti-His antibody is shown. ( E ) The expression of mFIZZ1 with wheat germ extract analysed on non-reducing 15% SDS-PAGE stained with Coomassie Brilliant Blue. ( F ) Strip of the respective immunoblot of ( E ) developed with anti-His antibody is shown. As marker (M) the PageRuler™ pre-stained Protein Ladder (Fermentas) is used. P = pellet, S = soluble fraction and T = total. The corresponding bands are indicated with an asterisk.
Bl21 De3, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "The Quiescin Sulfhydryl Oxidase (hQSOX1b) Tunes the Expression of Resistin-Like Molecule Alpha (RELM-? or mFIZZ1) in a Wheat Germ Cell-Free Extract"

Article Title: The Quiescin Sulfhydryl Oxidase (hQSOX1b) Tunes the Expression of Resistin-Like Molecule Alpha (RELM-? or mFIZZ1) in a Wheat Germ Cell-Free Extract

Journal: PLoS ONE

doi: 10.1371/journal.pone.0055621

mFIZZ1 soluble expressed using wheat germ extract. ( A ) The expression of mFIZZ1 in SHuffle™ T7, Origami DE3 and BL21 DE3 analysed on non-reducing 15% SDS-PAGE stained with Coomassie Brilliant Blue. ( B ) Strip of the immunoblot from the SDS-PAGE in ( A ) developed with anti-His antibody is shown. ( C ) The expression of mFIZZ1′ with wheat germ extract analysed on non-reducing 15% SDS-PAGE stained with Coomassie Brilliant Blue. ( D ) Strip of the respective immunoblot of ( C ) developed with anti-His antibody is shown. ( E ) The expression of mFIZZ1 with wheat germ extract analysed on non-reducing 15% SDS-PAGE stained with Coomassie Brilliant Blue. ( F ) Strip of the respective immunoblot of ( E ) developed with anti-His antibody is shown. As marker (M) the PageRuler™ pre-stained Protein Ladder (Fermentas) is used. P = pellet, S = soluble fraction and T = total. The corresponding bands are indicated with an asterisk.
Figure Legend Snippet: mFIZZ1 soluble expressed using wheat germ extract. ( A ) The expression of mFIZZ1 in SHuffle™ T7, Origami DE3 and BL21 DE3 analysed on non-reducing 15% SDS-PAGE stained with Coomassie Brilliant Blue. ( B ) Strip of the immunoblot from the SDS-PAGE in ( A ) developed with anti-His antibody is shown. ( C ) The expression of mFIZZ1′ with wheat germ extract analysed on non-reducing 15% SDS-PAGE stained with Coomassie Brilliant Blue. ( D ) Strip of the respective immunoblot of ( C ) developed with anti-His antibody is shown. ( E ) The expression of mFIZZ1 with wheat germ extract analysed on non-reducing 15% SDS-PAGE stained with Coomassie Brilliant Blue. ( F ) Strip of the respective immunoblot of ( E ) developed with anti-His antibody is shown. As marker (M) the PageRuler™ pre-stained Protein Ladder (Fermentas) is used. P = pellet, S = soluble fraction and T = total. The corresponding bands are indicated with an asterisk.

Techniques Used: Expressing, SDS Page, Staining, Stripping Membranes, Marker

2) Product Images from "A universal strategy for high-yield production of soluble and functional clostridial collagenases in E. coli"

Article Title: A universal strategy for high-yield production of soluble and functional clostridial collagenases in E. coli

Journal: Applied Microbiology and Biotechnology

doi: 10.1007/s00253-009-1953-4

Expression of ColG, H, and T constructs cloned in pET15b. For the nomenclature, compare Table 1 . M prestained protein ladder. Lane 1 BL21 DE3 cells before induction; lanes 2 – 5 hG1, hG2, hG3, and hG4 constructs of ColG 4 h after induction; lanes 6 – 9 hH1, hH2, hH3, and hH4 constructs of ColH 4 h after induction; lane 10 Tuner DE3 cells before induction; lanes 11 – 13 hT0, hT1, and hT3 constructs of ColT
Figure Legend Snippet: Expression of ColG, H, and T constructs cloned in pET15b. For the nomenclature, compare Table 1 . M prestained protein ladder. Lane 1 BL21 DE3 cells before induction; lanes 2 – 5 hG1, hG2, hG3, and hG4 constructs of ColG 4 h after induction; lanes 6 – 9 hH1, hH2, hH3, and hH4 constructs of ColH 4 h after induction; lane 10 Tuner DE3 cells before induction; lanes 11 – 13 hT0, hT1, and hT3 constructs of ColT

Techniques Used: Expressing, Construct, Clone Assay

Related Articles

Clone Assay:

Article Title: A simple vector system to improve performance and utilisation of recombinant antibodies
Article Snippet: Paragraph title: Reagents, scFv clones & bacterial strains ... E. coli BL21 (DE3) and Origami 2™ (DE3) cells and the pET26b(+) plasmid were from Novagen (San Diego, CA).

Article Title: Genomic and molecular characterization of a novel quorum sensing molecule in Bacillus licheniformis
Article Snippet: The expression strain [E. coli BL21 (DE3)] and E. coli TOP10 were used for cloning/transformations and were selected on LBA supplemented with ampicilin (100 µgml−1 ). .. E. coli BL21 ComX producer strain was cultivated in M9 minimal salts solution (sigma).

Article Title: The gaf Fimbrial Gene Cluster of Escherichia coli Expresses a Full-Size and a Truncated Soluble Adhesin Protein
Article Snippet: Plasmid pRR-5 contains the complete gaf gene cluster from E. coli strain IHE11165 on a 7-kb DNA fragment in pACYC184 , and pHUB110 contains a 6-bp in-frame deletion within the coding region of gafD , resulting in G fimbriae lacking GlcNAc-binding capacity ( ). pHUB113 ( ) contains the gafD reading frame cloned into pUC19. .. E. coli strain BL21 λDE3 ( ompT lon ) and expression vector pET-22b(+) were from Novagen Inc. (Madison, Wis.).

Centrifugation:

Article Title: A new hybrid bacteriocin, Ent35-MccV, displays antimicrobial activity against pathogenic Gram-positive and Gram-negative bacteria
Article Snippet: 2.3 Gene expression and bacteriocins purification for activity determinations Ent35–MccV was expressed from pMA24 in E. coli BL21 [DE3] (pLysS) grown in LB (Sigma) at 37 °C. .. After 3 h, cells were collected by centrifugation in 10 mM HEPES, pH 7, PMSF 0.5 mM (buffer A), lysed using a French press and centrifuged at 47,000xg , 4 °C for 1 h. The supernatant was heated (10 min, 100 °C) and centrifuged (10 min, 10,000xg ).

Article Title: Hypoallergenic Der p 1/Der p 2 combination vaccines for immunotherapy of house dust mite allergy
Article Snippet: Expression plasmids containing the Der p 2/1 constructs were transformed into E coli strain BL21 (DE3; Novagen, Merck). .. Protein expression was performed in 250 mL of liquid culture by means of induction with 0.5 mmol/L isopropyl-β-D-1-thiogalactopyranoside at an OD600 of 0.8 for 4 hours at 37°C, and cells were harvested by means of centrifugation at 4000 g for 15 minutes at 4°C.

Article Title: Purification and Reconstitution of the Connexin43 Carboxyl Terminus attached to the 4th Transmembrane Domain in Detergent Micelles *
Article Snippet: The E. coli strains BL21(DE3) (Novagen), Rosetta-2(DE3) (Novagen), C41(DE3) (Lucigen, ( )), and C43(DE3) (Lucigen, ( )) were transformed with the TM4-Cx43CT expression plasmid (see above) and then inoculated into 1 L of Luria Bertani medium (LB), enriched minimal media , or ISOGRO media (Sigma-Aldrich). .. The cells were harvested by centrifugation (1,000 × g for 45 min), washed with PBS buffer, and stored at −20°C.

Positron Emission Tomography:

Article Title: A Single Codon Optimization Enhances Recombinant Human TNF-α Vaccine Expression in Escherichia coli
Article Snippet: .. The vector pET-22b and E. coli strain BL21 (DE3) were purchased from Novagen (San Diego, CA). .. Isopropyl β -D-1-thiogalactopyranoside (IPTG) was from Sigma-Aldrich (St. Louis, MO).

Article Title: The gaf Fimbrial Gene Cluster of Escherichia coli Expresses a Full-Size and a Truncated Soluble Adhesin Protein
Article Snippet: .. E. coli strain BL21 λDE3 ( ompT lon ) and expression vector pET-22b(+) were from Novagen Inc. (Madison, Wis.). .. The bacteria were cultivated at 37°C in Luria broth ( ) containing the appropriate antibiotics.

Article Title: Identification and molecular characterization of a metagenome-derived L-lysine decarboxylase gene from subtropical soil microorganisms
Article Snippet: .. Plasmid pET-30a(+) (Novagen) and bacterial strain E . coli BL21 (DE3) pLysS (Novagen) were used as the expression vector and host, respectively. ..

Ligation:

Article Title: Chemical complexity of protein determines optimal E. coli expression host; A comparative study using Erythropoietin, Streptokinase and Tumor Necrosis Factor Receptor
Article Snippet: 2.1 Bacterial strains and plasmids For initial transformation of the ligation mixtures, the maintenance strain E. coli DH5α was used. .. E. coli BL21 (DE3), E. coli BL21 (DE3) pLys S, E. coli BL21 Rosetta (DE3) pLys S (Novagen,) are IPTG inducible strains in which the T7 RNA Polymerase gene is under the control of a Lac UV5 promoter.

Mouse Assay:

Article Title: A novel HPV16 E7-affitoxin for targeted therapy of HPV16-induced human cervical cancer
Article Snippet: ICR mice, weighing 23-27 g, were purchased from the animal experimental center of Wenzhou Medical University, China. .. The pET21a(+) vector and E.coli BL21 (DE3) were purchased from Novagen and ATCC, respectively.

Synthesized:

Article Title: Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of ADAMTS1 Inhibitors
Article Snippet: Materials E . coli BL21 (DE3), E. coli DH5a, pET32a(+) vector and his-bind purification kit were purchased from Novagen. .. The peptide, Abz-YPLPRNITEGEARGNVILTAK(Dnp)P-OH, was synthesized by GL Biochem Ltd. (Shanghai, China).

Article Title: Preparation and Antitumor Activity of CS5931, A Novel Polypeptide from Sea Squirt Ciona Savignyi
Article Snippet: .. Recombinant Expression of CS5931 from E. coli BL21 (DE3) The expression vector, pET28a/CS5931 synthesized by our lab previously [ ], was transfected into E. coli BL21 (DE3) cells (Novagen, Darmstadt, Germany). .. Cells were incubated in low-salt LB medium (containing 100 µg/mL Kanamycin) at 37 °C with shaking at 200 rpm.

Transfection:

Article Title: Preparation and Antitumor Activity of CS5931, A Novel Polypeptide from Sea Squirt Ciona Savignyi
Article Snippet: .. Recombinant Expression of CS5931 from E. coli BL21 (DE3) The expression vector, pET28a/CS5931 synthesized by our lab previously [ ], was transfected into E. coli BL21 (DE3) cells (Novagen, Darmstadt, Germany). .. Cells were incubated in low-salt LB medium (containing 100 µg/mL Kanamycin) at 37 °C with shaking at 200 rpm.

Construct:

Article Title: Hypoallergenic Der p 1/Der p 2 combination vaccines for immunotherapy of house dust mite allergy
Article Snippet: .. Expression plasmids containing the Der p 2/1 constructs were transformed into E coli strain BL21 (DE3; Novagen, Merck). .. Protein expression was performed in 250 mL of liquid culture by means of induction with 0.5 mmol/L isopropyl-β-D-1-thiogalactopyranoside at an OD600 of 0.8 for 4 hours at 37°C, and cells were harvested by means of centrifugation at 4000 g for 15 minutes at 4°C.

Purification:

Article Title: A new hybrid bacteriocin, Ent35-MccV, displays antimicrobial activity against pathogenic Gram-positive and Gram-negative bacteria
Article Snippet: .. 2.3 Gene expression and bacteriocins purification for activity determinations Ent35–MccV was expressed from pMA24 in E. coli BL21 [DE3] (pLysS) grown in LB (Sigma) at 37 °C. ..

Article Title: Hypoallergenic Der p 1/Der p 2 combination vaccines for immunotherapy of house dust mite allergy
Article Snippet: Paragraph title: Expression and purification of Der p 2/1 mosaic proteins ... Expression plasmids containing the Der p 2/1 constructs were transformed into E coli strain BL21 (DE3; Novagen, Merck).

Article Title: Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of ADAMTS1 Inhibitors
Article Snippet: .. Materials E . coli BL21 (DE3), E. coli DH5a, pET32a(+) vector and his-bind purification kit were purchased from Novagen. .. Heparin sepharose CL-6B column was purchased from Amersham Biosciences.

Article Title: Purification and Reconstitution of the Connexin43 Carboxyl Terminus attached to the 4th Transmembrane Domain in Detergent Micelles *
Article Snippet: Paragraph title: Protein expression and purification ... The E. coli strains BL21(DE3) (Novagen), Rosetta-2(DE3) (Novagen), C41(DE3) (Lucigen, ( )), and C43(DE3) (Lucigen, ( )) were transformed with the TM4-Cx43CT expression plasmid (see above) and then inoculated into 1 L of Luria Bertani medium (LB), enriched minimal media , or ISOGRO media (Sigma-Aldrich).

Plasmid Preparation:

Article Title: A simple vector system to improve performance and utilisation of recombinant antibodies
Article Snippet: .. E. coli BL21 (DE3) and Origami 2™ (DE3) cells and the pET26b(+) plasmid were from Novagen (San Diego, CA). ..

Article Title: Chemical complexity of protein determines optimal E. coli expression host; A comparative study using Erythropoietin, Streptokinase and Tumor Necrosis Factor Receptor
Article Snippet: E. coli BL21 (DE3), E. coli BL21 (DE3) pLys S, E. coli BL21 Rosetta (DE3) pLys S (Novagen,) are IPTG inducible strains in which the T7 RNA Polymerase gene is under the control of a Lac UV5 promoter. .. The expression vector used in this study is pRSET A obtained from Invitrogen Life Technologies, USA. pRSET is a high copy number plasmid with a pUC origin of replication.

Article Title: A Single Codon Optimization Enhances Recombinant Human TNF-α Vaccine Expression in Escherichia coli
Article Snippet: .. The vector pET-22b and E. coli strain BL21 (DE3) were purchased from Novagen (San Diego, CA). .. Isopropyl β -D-1-thiogalactopyranoside (IPTG) was from Sigma-Aldrich (St. Louis, MO).

Article Title: The gaf Fimbrial Gene Cluster of Escherichia coli Expresses a Full-Size and a Truncated Soluble Adhesin Protein
Article Snippet: .. E. coli strain BL21 λDE3 ( ompT lon ) and expression vector pET-22b(+) were from Novagen Inc. (Madison, Wis.). .. The bacteria were cultivated at 37°C in Luria broth ( ) containing the appropriate antibiotics.

Article Title: A novel HPV16 E7-affitoxin for targeted therapy of HPV16-induced human cervical cancer
Article Snippet: .. The pET21a(+) vector and E.coli BL21 (DE3) were purchased from Novagen and ATCC, respectively. .. Reagents The reagents used, including Cell Counting Kit-8 (CCK-8) (Dojindo, Japan), RPMI-1640 (Gibco, USA), fetal bovine serum (FBS) (Gibco, USA), penicillin (Gibco, USA), trypsin-EDTA (Gibco, USA), streptomycin (Sigma Aldrich, Saint Louis, USA), Isopropyl-D-thiogalactopyranoside (IPTG) (Sigma Aldrich, Saint Louis, USA), Ni-NTA agarose (Qiagen Inc., Valencia, CA), and DyLight-755 (Thermo Fisher Scientific, USA), were purchased from commercial sources.

Article Title: Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of ADAMTS1 Inhibitors
Article Snippet: .. Materials E . coli BL21 (DE3), E. coli DH5a, pET32a(+) vector and his-bind purification kit were purchased from Novagen. .. Heparin sepharose CL-6B column was purchased from Amersham Biosciences.

Article Title: Preparation and Antitumor Activity of CS5931, A Novel Polypeptide from Sea Squirt Ciona Savignyi
Article Snippet: .. Recombinant Expression of CS5931 from E. coli BL21 (DE3) The expression vector, pET28a/CS5931 synthesized by our lab previously [ ], was transfected into E. coli BL21 (DE3) cells (Novagen, Darmstadt, Germany). .. Cells were incubated in low-salt LB medium (containing 100 µg/mL Kanamycin) at 37 °C with shaking at 200 rpm.

Article Title: Identification and molecular characterization of a metagenome-derived L-lysine decarboxylase gene from subtropical soil microorganisms
Article Snippet: .. Plasmid pET-30a(+) (Novagen) and bacterial strain E . coli BL21 (DE3) pLysS (Novagen) were used as the expression vector and host, respectively. ..

Article Title: Purification and Reconstitution of the Connexin43 Carboxyl Terminus attached to the 4th Transmembrane Domain in Detergent Micelles *
Article Snippet: .. The E. coli strains BL21(DE3) (Novagen), Rosetta-2(DE3) (Novagen), C41(DE3) (Lucigen, ( )), and C43(DE3) (Lucigen, ( )) were transformed with the TM4-Cx43CT expression plasmid (see above) and then inoculated into 1 L of Luria Bertani medium (LB), enriched minimal media , or ISOGRO media (Sigma-Aldrich). .. Cultures were incubated at 37°C with continuous agitation.

Concentration Assay:

Article Title: Preparation and Antitumor Activity of CS5931, A Novel Polypeptide from Sea Squirt Ciona Savignyi
Article Snippet: Recombinant Expression of CS5931 from E. coli BL21 (DE3) The expression vector, pET28a/CS5931 synthesized by our lab previously [ ], was transfected into E. coli BL21 (DE3) cells (Novagen, Darmstadt, Germany). .. After the value of OD600 of the culture medium reached around 0.5–0.7, IPTG was added at a final concentration of 0.75 mM and the cells were incubated overnight at 16 °C.

Incubation:

Article Title: Hypoallergenic Der p 1/Der p 2 combination vaccines for immunotherapy of house dust mite allergy
Article Snippet: Expression plasmids containing the Der p 2/1 constructs were transformed into E coli strain BL21 (DE3; Novagen, Merck). .. Cells were lysed by using 3 freeze/thaw cycles (−70°C/+50°C), DNA was degraded by means of incubation with 1 μg of DNase I for 10 minutes at room temperature, and cell debris was removed by means of centrifugation (10,000 g for 30 minutes at 4°C).

Article Title: Preparation and Antitumor Activity of CS5931, A Novel Polypeptide from Sea Squirt Ciona Savignyi
Article Snippet: Recombinant Expression of CS5931 from E. coli BL21 (DE3) The expression vector, pET28a/CS5931 synthesized by our lab previously [ ], was transfected into E. coli BL21 (DE3) cells (Novagen, Darmstadt, Germany). .. Cells were incubated in low-salt LB medium (containing 100 µg/mL Kanamycin) at 37 °C with shaking at 200 rpm.

Article Title: Purification and Reconstitution of the Connexin43 Carboxyl Terminus attached to the 4th Transmembrane Domain in Detergent Micelles *
Article Snippet: The E. coli strains BL21(DE3) (Novagen), Rosetta-2(DE3) (Novagen), C41(DE3) (Lucigen, ( )), and C43(DE3) (Lucigen, ( )) were transformed with the TM4-Cx43CT expression plasmid (see above) and then inoculated into 1 L of Luria Bertani medium (LB), enriched minimal media , or ISOGRO media (Sigma-Aldrich). .. Cultures were incubated at 37°C with continuous agitation.

Cell Culture:

Article Title: A novel HPV16 E7-affitoxin for targeted therapy of HPV16-induced human cervical cancer
Article Snippet: SiHa (ATCC: HTB-35, HPV16 positive, contains about one to two copies of integrated HPV16 genome), CaSki (ATCC: CRL-1550, HPV16 positive, contains about 600 copies of integrated HPV16 genome), HeLa 229 (ATCC: CCL-2.1, HPV18 positive, used as HPV16 negative control cell line), and melanoma tumor A375 (ATCC: CRL-1619, used as HPV negative control cell line) were obtained from the American Type Culture Collection (ATCC, USA) and cultured as previously described . .. The pET21a(+) vector and E.coli BL21 (DE3) were purchased from Novagen and ATCC, respectively.

Activity Assay:

Article Title: A new hybrid bacteriocin, Ent35-MccV, displays antimicrobial activity against pathogenic Gram-positive and Gram-negative bacteria
Article Snippet: .. 2.3 Gene expression and bacteriocins purification for activity determinations Ent35–MccV was expressed from pMA24 in E. coli BL21 [DE3] (pLysS) grown in LB (Sigma) at 37 °C. ..

Negative Control:

Article Title: A novel HPV16 E7-affitoxin for targeted therapy of HPV16-induced human cervical cancer
Article Snippet: SiHa (ATCC: HTB-35, HPV16 positive, contains about one to two copies of integrated HPV16 genome), CaSki (ATCC: CRL-1550, HPV16 positive, contains about 600 copies of integrated HPV16 genome), HeLa 229 (ATCC: CCL-2.1, HPV18 positive, used as HPV16 negative control cell line), and melanoma tumor A375 (ATCC: CRL-1619, used as HPV negative control cell line) were obtained from the American Type Culture Collection (ATCC, USA) and cultured as previously described . .. The pET21a(+) vector and E.coli BL21 (DE3) were purchased from Novagen and ATCC, respectively.

Expressing:

Article Title: A new hybrid bacteriocin, Ent35-MccV, displays antimicrobial activity against pathogenic Gram-positive and Gram-negative bacteria
Article Snippet: .. 2.3 Gene expression and bacteriocins purification for activity determinations Ent35–MccV was expressed from pMA24 in E. coli BL21 [DE3] (pLysS) grown in LB (Sigma) at 37 °C. ..

Article Title: Hypoallergenic Der p 1/Der p 2 combination vaccines for immunotherapy of house dust mite allergy
Article Snippet: .. Expression plasmids containing the Der p 2/1 constructs were transformed into E coli strain BL21 (DE3; Novagen, Merck). .. Protein expression was performed in 250 mL of liquid culture by means of induction with 0.5 mmol/L isopropyl-β-D-1-thiogalactopyranoside at an OD600 of 0.8 for 4 hours at 37°C, and cells were harvested by means of centrifugation at 4000 g for 15 minutes at 4°C.

Article Title: Chemical complexity of protein determines optimal E. coli expression host; A comparative study using Erythropoietin, Streptokinase and Tumor Necrosis Factor Receptor
Article Snippet: The bacterial strains used to study expression were E. coli GJ1158, E. coli BL21 (DE3), E. coli BL21 (DE3) pLys S, E. coli BL21 Rosetta (DE3) pLys S. E. coli GJ1158 , derived from E. coli B strain BL21, is a salt inducible strain with a pro U promoter ( ). .. E. coli BL21 (DE3), E. coli BL21 (DE3) pLys S, E. coli BL21 Rosetta (DE3) pLys S (Novagen,) are IPTG inducible strains in which the T7 RNA Polymerase gene is under the control of a Lac UV5 promoter.

Article Title: Genomic and molecular characterization of a novel quorum sensing molecule in Bacillus licheniformis
Article Snippet: The expression strain [E. coli BL21 (DE3)] and E. coli TOP10 were used for cloning/transformations and were selected on LBA supplemented with ampicilin (100 µgml−1 ). .. E. coli BL21 ComX producer strain was cultivated in M9 minimal salts solution (sigma).

Article Title: The gaf Fimbrial Gene Cluster of Escherichia coli Expresses a Full-Size and a Truncated Soluble Adhesin Protein
Article Snippet: .. E. coli strain BL21 λDE3 ( ompT lon ) and expression vector pET-22b(+) were from Novagen Inc. (Madison, Wis.). .. The bacteria were cultivated at 37°C in Luria broth ( ) containing the appropriate antibiotics.

Article Title: Preparation and Antitumor Activity of CS5931, A Novel Polypeptide from Sea Squirt Ciona Savignyi
Article Snippet: .. Recombinant Expression of CS5931 from E. coli BL21 (DE3) The expression vector, pET28a/CS5931 synthesized by our lab previously [ ], was transfected into E. coli BL21 (DE3) cells (Novagen, Darmstadt, Germany). .. Cells were incubated in low-salt LB medium (containing 100 µg/mL Kanamycin) at 37 °C with shaking at 200 rpm.

Article Title: Purification and Reconstitution of the Connexin43 Carboxyl Terminus attached to the 4th Transmembrane Domain in Detergent Micelles *
Article Snippet: .. The E. coli strains BL21(DE3) (Novagen), Rosetta-2(DE3) (Novagen), C41(DE3) (Lucigen, ( )), and C43(DE3) (Lucigen, ( )) were transformed with the TM4-Cx43CT expression plasmid (see above) and then inoculated into 1 L of Luria Bertani medium (LB), enriched minimal media , or ISOGRO media (Sigma-Aldrich). .. Cultures were incubated at 37°C with continuous agitation.

Transformation Assay:

Article Title: Hypoallergenic Der p 1/Der p 2 combination vaccines for immunotherapy of house dust mite allergy
Article Snippet: .. Expression plasmids containing the Der p 2/1 constructs were transformed into E coli strain BL21 (DE3; Novagen, Merck). .. Protein expression was performed in 250 mL of liquid culture by means of induction with 0.5 mmol/L isopropyl-β-D-1-thiogalactopyranoside at an OD600 of 0.8 for 4 hours at 37°C, and cells were harvested by means of centrifugation at 4000 g for 15 minutes at 4°C.

Article Title: Chemical complexity of protein determines optimal E. coli expression host; A comparative study using Erythropoietin, Streptokinase and Tumor Necrosis Factor Receptor
Article Snippet: 2.1 Bacterial strains and plasmids For initial transformation of the ligation mixtures, the maintenance strain E. coli DH5α was used. .. E. coli BL21 (DE3), E. coli BL21 (DE3) pLys S, E. coli BL21 Rosetta (DE3) pLys S (Novagen,) are IPTG inducible strains in which the T7 RNA Polymerase gene is under the control of a Lac UV5 promoter.

Article Title: Purification and Reconstitution of the Connexin43 Carboxyl Terminus attached to the 4th Transmembrane Domain in Detergent Micelles *
Article Snippet: .. The E. coli strains BL21(DE3) (Novagen), Rosetta-2(DE3) (Novagen), C41(DE3) (Lucigen, ( )), and C43(DE3) (Lucigen, ( )) were transformed with the TM4-Cx43CT expression plasmid (see above) and then inoculated into 1 L of Luria Bertani medium (LB), enriched minimal media , or ISOGRO media (Sigma-Aldrich). .. Cultures were incubated at 37°C with continuous agitation.

Recombinant:

Article Title: Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of ADAMTS1 Inhibitors
Article Snippet: Materials E . coli BL21 (DE3), E. coli DH5a, pET32a(+) vector and his-bind purification kit were purchased from Novagen. .. Recombinant human ADAMTS1 was purchased from R & D Systems.

Article Title: Preparation and Antitumor Activity of CS5931, A Novel Polypeptide from Sea Squirt Ciona Savignyi
Article Snippet: .. Recombinant Expression of CS5931 from E. coli BL21 (DE3) The expression vector, pET28a/CS5931 synthesized by our lab previously [ ], was transfected into E. coli BL21 (DE3) cells (Novagen, Darmstadt, Germany). .. Cells were incubated in low-salt LB medium (containing 100 µg/mL Kanamycin) at 37 °C with shaking at 200 rpm.

Derivative Assay:

Article Title: Chemical complexity of protein determines optimal E. coli expression host; A comparative study using Erythropoietin, Streptokinase and Tumor Necrosis Factor Receptor
Article Snippet: The bacterial strains used to study expression were E. coli GJ1158, E. coli BL21 (DE3), E. coli BL21 (DE3) pLys S, E. coli BL21 Rosetta (DE3) pLys S. E. coli GJ1158 , derived from E. coli B strain BL21, is a salt inducible strain with a pro U promoter ( ). .. E. coli BL21 (DE3), E. coli BL21 (DE3) pLys S, E. coli BL21 Rosetta (DE3) pLys S (Novagen,) are IPTG inducible strains in which the T7 RNA Polymerase gene is under the control of a Lac UV5 promoter.

Generated:

Article Title: A simple vector system to improve performance and utilisation of recombinant antibodies
Article Snippet: E. coli BL21 (DE3) and Origami 2™ (DE3) cells and the pET26b(+) plasmid were from Novagen (San Diego, CA). .. The other scFvs were generated in house to commercially available antigens purchased from R & D Systems (MN, USA), USBiologicals (MA, USA) and Progen (Heidelberg, Germany).

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    Millipore e coli strain bl21
    Zymographic detection of the autolytic activity in recombinant His-tagged Atl. Protein extracts from E. coli <t>BL21</t> cells grown with IPTG were used in this study. The protein extract was separated by 12.5% SDS-gel electrophoresis. The SDS gel was impregnated with autoclaved S. aureus RN450 cells. Autolytic activity was detected by renaturing the autolysins by incubation of the gel in 50 mM Tri-HCl buffer, pH 7.5, containing 0.05% Triton X-100. Lane 1: full length His-tagged Atl; lane 2: Atl-1; lane 3: Atl-2; lane 4: Atl-3; lane 5: Atl-4; lane 6: Atl-5; lane 7: Atl-6; lane 8: Atl-7; lane 9: Atl-8.
    E Coli Strain Bl21, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 268 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore lsrb bl21 ∆ luxs protein
    Binding of <t>LsrB</t> to endogenous AI-2. Purified LsrB (BL21) and LsrB <t>(BL21∆luxS)</t> proteins (5 mg/ml) were incubated for 10 min at 37, 50 and 60 °C to release endogenous AI-2, respectively. After incubation, the LsrB proteins were removed by ultrafiltration (10,000-Da cut-off; EMD Millipore), and the filtered reaction products were tested for AI-2 activity using a V. harveyi BB170 bioassay. The extent of LsrB binding to endogenous AI-2 was evaluated using an AI-2 assay, which showed that recombinant LsrB (BL21) bound to endogenous AI-2 (produced by wild-type strain BL21) and was released from LsrB (BL21) at 50 or 60 °C, respectively ( a ). However, since the luxS mutant BL21∆luxS did not produce endogenous AI-2, no AI-2 could be released from the recombinant LsrB (BL21∆luxS) ( b ). Moreover, the recombinant LuxS protein, which was expressed in strain BL21 (pColdTF-lsrB) as a negative control, also showed no AI-2 binding activity ( c ). AI-2 (10 μM) was used as a positive control
    Lsrb Bl21 ∆ Luxs Protein, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore bl21 ∆ luxs
    Construction and identification of luxS mutant strain <t>BL21∆luxS</t>
    Bl21 ∆ Luxs, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 ∆ luxs/product/Millipore
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    86
    Millipore buffer a bl21 pes2i pellets
    SDS-PAGE analysis of purified protein . Shown are the CaroS2K (A) and CaroS2I (B). Samples were subjected to electrophoresis in 10% polyacrylamide gels, which were stained with Coomassie blue. Lane M, molecular weight standards (kDa); lane 1, cell lysate of E. coli <t>BL21/pET32a;</t> lane 4, cell lysate of BL21/pET30b; lanes 2 and 5, IPTG-induced cell lysates of BL21/pES2kI and <t>BL21/pES2I,</t> respectively; lanes 3 and 6, purified protein obtained after elution. The arrowheads indicate the killing protein of carocin S2K (A) and the immunity protein of carocin S2I (B).
    Buffer A Bl21 Pes2i Pellets, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Zymographic detection of the autolytic activity in recombinant His-tagged Atl. Protein extracts from E. coli BL21 cells grown with IPTG were used in this study. The protein extract was separated by 12.5% SDS-gel electrophoresis. The SDS gel was impregnated with autoclaved S. aureus RN450 cells. Autolytic activity was detected by renaturing the autolysins by incubation of the gel in 50 mM Tri-HCl buffer, pH 7.5, containing 0.05% Triton X-100. Lane 1: full length His-tagged Atl; lane 2: Atl-1; lane 3: Atl-2; lane 4: Atl-3; lane 5: Atl-4; lane 6: Atl-5; lane 7: Atl-6; lane 8: Atl-7; lane 9: Atl-8.

    Journal: International Journal of Microbiology

    Article Title: High Level Expression and Purification of Atl, the Major Autolytic Protein of Staphylococcus aureus

    doi: 10.1155/2014/615965

    Figure Lengend Snippet: Zymographic detection of the autolytic activity in recombinant His-tagged Atl. Protein extracts from E. coli BL21 cells grown with IPTG were used in this study. The protein extract was separated by 12.5% SDS-gel electrophoresis. The SDS gel was impregnated with autoclaved S. aureus RN450 cells. Autolytic activity was detected by renaturing the autolysins by incubation of the gel in 50 mM Tri-HCl buffer, pH 7.5, containing 0.05% Triton X-100. Lane 1: full length His-tagged Atl; lane 2: Atl-1; lane 3: Atl-2; lane 4: Atl-3; lane 5: Atl-4; lane 6: Atl-5; lane 7: Atl-6; lane 8: Atl-7; lane 9: Atl-8.

    Article Snippet: In addition, E. coli strain BL21 (EMD Millipore) was used for all protein expression studies.

    Techniques: Activity Assay, Recombinant, SDS-Gel, Electrophoresis, Incubation

    Coomassie stained gels showing purity of recombinant His-tagged Atl. Recombinant Atl and Atl-1 proteins were overproduced in E. coli and purified as described in the Materials and Methods section. Lane M: standard protein markers; lanes 1 and 2: protein extract from E. coli BL21 cells transformed with plasmids pRSETA- atl and grown without and with IPTG, respectively; lane 3: purified His-tagged Atl; lanes 4 and 5: protein extract from E. coli BL21 cells transformed with plasmids pRSETA- atl1 and grown without and with IPTG, respectively; lane 6: purified His-tagged Atl-1.

    Journal: International Journal of Microbiology

    Article Title: High Level Expression and Purification of Atl, the Major Autolytic Protein of Staphylococcus aureus

    doi: 10.1155/2014/615965

    Figure Lengend Snippet: Coomassie stained gels showing purity of recombinant His-tagged Atl. Recombinant Atl and Atl-1 proteins were overproduced in E. coli and purified as described in the Materials and Methods section. Lane M: standard protein markers; lanes 1 and 2: protein extract from E. coli BL21 cells transformed with plasmids pRSETA- atl and grown without and with IPTG, respectively; lane 3: purified His-tagged Atl; lanes 4 and 5: protein extract from E. coli BL21 cells transformed with plasmids pRSETA- atl1 and grown without and with IPTG, respectively; lane 6: purified His-tagged Atl-1.

    Article Snippet: In addition, E. coli strain BL21 (EMD Millipore) was used for all protein expression studies.

    Techniques: Staining, Recombinant, Purification, Transformation Assay

    Zymographic detection of the autolytic activity in purified recombinant His-tagged Atl. Lane 1: total protein extract from S. aureus cells. Lanes 2 and 4: purified His-tagged Atl and Atl-1, respectively; lanes 3 and 5: protein extract from E. coli BL21 cells transformed with plasmids pRSETA- atl or pRSETA- atl1 and grown with IPTG, respectively.

    Journal: International Journal of Microbiology

    Article Title: High Level Expression and Purification of Atl, the Major Autolytic Protein of Staphylococcus aureus

    doi: 10.1155/2014/615965

    Figure Lengend Snippet: Zymographic detection of the autolytic activity in purified recombinant His-tagged Atl. Lane 1: total protein extract from S. aureus cells. Lanes 2 and 4: purified His-tagged Atl and Atl-1, respectively; lanes 3 and 5: protein extract from E. coli BL21 cells transformed with plasmids pRSETA- atl or pRSETA- atl1 and grown with IPTG, respectively.

    Article Snippet: In addition, E. coli strain BL21 (EMD Millipore) was used for all protein expression studies.

    Techniques: Activity Assay, Purification, Recombinant, Transformation Assay

    Coomassie stained gels demonstrating overexpression of recombinant Atl. The 12.5% SDS-PAGE contains protein extracts of E. coli BL21 cells with pRSETA plasmids expressing either full length or truncated Atl proteins. The odd number labels are cells grown without IPTG and the even number labels are the cells grown with IPTG. Lane M: standard protein markers; lanes 1 and 2 - Atl (full length His-tagged Atl); lanes 3 and 4: Atl-1; lanes 5 and 6: Atl-2; lanes 7 and 8: Atl-3; lanes 9 and 10: Atl-4; lanes 11 and 12: Atl-6; lanes 13 and 14: Atl-5; lanes 15 and 16: Atl-7; lanes 17 and 18: Atl-8. The Atl number suffixes are indicated in Table 1 .

    Journal: International Journal of Microbiology

    Article Title: High Level Expression and Purification of Atl, the Major Autolytic Protein of Staphylococcus aureus

    doi: 10.1155/2014/615965

    Figure Lengend Snippet: Coomassie stained gels demonstrating overexpression of recombinant Atl. The 12.5% SDS-PAGE contains protein extracts of E. coli BL21 cells with pRSETA plasmids expressing either full length or truncated Atl proteins. The odd number labels are cells grown without IPTG and the even number labels are the cells grown with IPTG. Lane M: standard protein markers; lanes 1 and 2 - Atl (full length His-tagged Atl); lanes 3 and 4: Atl-1; lanes 5 and 6: Atl-2; lanes 7 and 8: Atl-3; lanes 9 and 10: Atl-4; lanes 11 and 12: Atl-6; lanes 13 and 14: Atl-5; lanes 15 and 16: Atl-7; lanes 17 and 18: Atl-8. The Atl number suffixes are indicated in Table 1 .

    Article Snippet: In addition, E. coli strain BL21 (EMD Millipore) was used for all protein expression studies.

    Techniques: Staining, Over Expression, Recombinant, SDS Page, Expressing

    Binding of LsrB to endogenous AI-2. Purified LsrB (BL21) and LsrB (BL21∆luxS) proteins (5 mg/ml) were incubated for 10 min at 37, 50 and 60 °C to release endogenous AI-2, respectively. After incubation, the LsrB proteins were removed by ultrafiltration (10,000-Da cut-off; EMD Millipore), and the filtered reaction products were tested for AI-2 activity using a V. harveyi BB170 bioassay. The extent of LsrB binding to endogenous AI-2 was evaluated using an AI-2 assay, which showed that recombinant LsrB (BL21) bound to endogenous AI-2 (produced by wild-type strain BL21) and was released from LsrB (BL21) at 50 or 60 °C, respectively ( a ). However, since the luxS mutant BL21∆luxS did not produce endogenous AI-2, no AI-2 could be released from the recombinant LsrB (BL21∆luxS) ( b ). Moreover, the recombinant LuxS protein, which was expressed in strain BL21 (pColdTF-lsrB) as a negative control, also showed no AI-2 binding activity ( c ). AI-2 (10 μM) was used as a positive control

    Journal: AMB Express

    Article Title: LsrB-based and temperature-dependent identification of bacterial AI-2 receptor

    doi: 10.1186/s13568-017-0486-y

    Figure Lengend Snippet: Binding of LsrB to endogenous AI-2. Purified LsrB (BL21) and LsrB (BL21∆luxS) proteins (5 mg/ml) were incubated for 10 min at 37, 50 and 60 °C to release endogenous AI-2, respectively. After incubation, the LsrB proteins were removed by ultrafiltration (10,000-Da cut-off; EMD Millipore), and the filtered reaction products were tested for AI-2 activity using a V. harveyi BB170 bioassay. The extent of LsrB binding to endogenous AI-2 was evaluated using an AI-2 assay, which showed that recombinant LsrB (BL21) bound to endogenous AI-2 (produced by wild-type strain BL21) and was released from LsrB (BL21) at 50 or 60 °C, respectively ( a ). However, since the luxS mutant BL21∆luxS did not produce endogenous AI-2, no AI-2 could be released from the recombinant LsrB (BL21∆luxS) ( b ). Moreover, the recombinant LuxS protein, which was expressed in strain BL21 (pColdTF-lsrB) as a negative control, also showed no AI-2 binding activity ( c ). AI-2 (10 μM) was used as a positive control

    Article Snippet: After incubation, the LsrB (BL21∆luxS) protein was removed by ultrafiltration (10,000-Da cut-off, EMD Millipore) and the filtered reaction products were tested for AI-2 activity by investigating the binding activity of LsrB (BL21∆luxS) to exogenous AI-2.

    Techniques: Binding Assay, Purification, Incubation, Activity Assay, Recombinant, Produced, Mutagenesis, Negative Control, Positive Control

    Schematic chart of strategy for binding of LsrB to endogenous AI-2 or exogenous AI-2. The extent of LsrB binding to endogenous AI-2 was evaluated using an AI-2 assay, which showed that recombinant LsrB (BL21) bound to endogenous AI-2 (produced by wild-type strain BL21) and was released from LsrB (BL21) at 50 or 60 °C. However, since the luxS mutant BL21∆luxS did not produce endogenous AI-2, no AI-2 could be released from the recombinant LsrB (BL21∆luxS). The combination of endogenous AI-2 produced by BL21 (DE3), which is known to interfere with the binding of AI-2 to LsrB (BL21), resulted in the loss in the ability of LsrB (BL21) to bind to exogenous AI-2. Hence, the BL21 (DE3) luxS mutant, which was incapable of producing endogenous AI-2, but could bind to exogenous AI-2

    Journal: AMB Express

    Article Title: LsrB-based and temperature-dependent identification of bacterial AI-2 receptor

    doi: 10.1186/s13568-017-0486-y

    Figure Lengend Snippet: Schematic chart of strategy for binding of LsrB to endogenous AI-2 or exogenous AI-2. The extent of LsrB binding to endogenous AI-2 was evaluated using an AI-2 assay, which showed that recombinant LsrB (BL21) bound to endogenous AI-2 (produced by wild-type strain BL21) and was released from LsrB (BL21) at 50 or 60 °C. However, since the luxS mutant BL21∆luxS did not produce endogenous AI-2, no AI-2 could be released from the recombinant LsrB (BL21∆luxS). The combination of endogenous AI-2 produced by BL21 (DE3), which is known to interfere with the binding of AI-2 to LsrB (BL21), resulted in the loss in the ability of LsrB (BL21) to bind to exogenous AI-2. Hence, the BL21 (DE3) luxS mutant, which was incapable of producing endogenous AI-2, but could bind to exogenous AI-2

    Article Snippet: After incubation, the LsrB (BL21∆luxS) protein was removed by ultrafiltration (10,000-Da cut-off, EMD Millipore) and the filtered reaction products were tested for AI-2 activity by investigating the binding activity of LsrB (BL21∆luxS) to exogenous AI-2.

    Techniques: Binding Assay, Recombinant, Produced, Mutagenesis

    SDS-PAGE analysis of total cellular proteins and purified fusion proteins from BL21∆luxS cells. Lane 1 and lane 5: SDS-PAGE analysis of total cellular proteins from BL21∆luxS containing pCold TF (serve as negative control). Lane 2 and lane 6: SDS-PAGE analysis of total cellular proteins containing expression plasmids pCold-TF-lsrB ( a ) and pCold-TF-luxS ( b ), respectively. Lane 3 and lane 7: SDS-PAGE analysis of total precipitation proteins containing expression plasmids pCold-TF-lsrB and pCold-TF-luxS, respectively. Lane 4 and lane 8: elution of the LsrB and LuxS purified fusion protein from the affinity column, respectively

    Journal: AMB Express

    Article Title: LsrB-based and temperature-dependent identification of bacterial AI-2 receptor

    doi: 10.1186/s13568-017-0486-y

    Figure Lengend Snippet: SDS-PAGE analysis of total cellular proteins and purified fusion proteins from BL21∆luxS cells. Lane 1 and lane 5: SDS-PAGE analysis of total cellular proteins from BL21∆luxS containing pCold TF (serve as negative control). Lane 2 and lane 6: SDS-PAGE analysis of total cellular proteins containing expression plasmids pCold-TF-lsrB ( a ) and pCold-TF-luxS ( b ), respectively. Lane 3 and lane 7: SDS-PAGE analysis of total precipitation proteins containing expression plasmids pCold-TF-lsrB and pCold-TF-luxS, respectively. Lane 4 and lane 8: elution of the LsrB and LuxS purified fusion protein from the affinity column, respectively

    Article Snippet: After incubation, the LsrB (BL21∆luxS) protein was removed by ultrafiltration (10,000-Da cut-off, EMD Millipore) and the filtered reaction products were tested for AI-2 activity by investigating the binding activity of LsrB (BL21∆luxS) to exogenous AI-2.

    Techniques: SDS Page, Purification, Negative Control, Expressing, Affinity Column

    Construction and identification of luxS mutant strain BL21∆luxS

    Journal: AMB Express

    Article Title: LsrB-based and temperature-dependent identification of bacterial AI-2 receptor

    doi: 10.1186/s13568-017-0486-y

    Figure Lengend Snippet: Construction and identification of luxS mutant strain BL21∆luxS

    Article Snippet: Cell-free culture fluid (CF) was prepared as follows: E. coli strains BL21 (DE3), BL21∆luxS and DH5a were grown in LB at 37 °C, and then pelleted by centrifugation at 12,000 g at 4 °C for 10 min. Then, the resulting supernatants were filtered through a 0.22-μm filter (EMD Millipore, Bedford, MA, USA) to obtain CF samples.

    Techniques: Mutagenesis

    The AI-2 activity in BL21∆luxS. The wild strain BL21 secretes AI-2-like molecules, and can induce V . harveyi BB170 bioluminescence, whereas no bioluminescence induction was observed for the mutant BL21∆ luxS . V . harveyi BB152 served as

    Journal: AMB Express

    Article Title: LsrB-based and temperature-dependent identification of bacterial AI-2 receptor

    doi: 10.1186/s13568-017-0486-y

    Figure Lengend Snippet: The AI-2 activity in BL21∆luxS. The wild strain BL21 secretes AI-2-like molecules, and can induce V . harveyi BB170 bioluminescence, whereas no bioluminescence induction was observed for the mutant BL21∆ luxS . V . harveyi BB152 served as

    Article Snippet: Cell-free culture fluid (CF) was prepared as follows: E. coli strains BL21 (DE3), BL21∆luxS and DH5a were grown in LB at 37 °C, and then pelleted by centrifugation at 12,000 g at 4 °C for 10 min. Then, the resulting supernatants were filtered through a 0.22-μm filter (EMD Millipore, Bedford, MA, USA) to obtain CF samples.

    Techniques: Activity Assay, Mutagenesis

    SDS-PAGE analysis of total cellular proteins and purified fusion proteins from BL21∆luxS cells. Lane 1 and lane 5: SDS-PAGE analysis of total cellular proteins from BL21∆luxS containing pCold TF (serve as negative control). Lane 2 and

    Journal: AMB Express

    Article Title: LsrB-based and temperature-dependent identification of bacterial AI-2 receptor

    doi: 10.1186/s13568-017-0486-y

    Figure Lengend Snippet: SDS-PAGE analysis of total cellular proteins and purified fusion proteins from BL21∆luxS cells. Lane 1 and lane 5: SDS-PAGE analysis of total cellular proteins from BL21∆luxS containing pCold TF (serve as negative control). Lane 2 and

    Article Snippet: Cell-free culture fluid (CF) was prepared as follows: E. coli strains BL21 (DE3), BL21∆luxS and DH5a were grown in LB at 37 °C, and then pelleted by centrifugation at 12,000 g at 4 °C for 10 min. Then, the resulting supernatants were filtered through a 0.22-μm filter (EMD Millipore, Bedford, MA, USA) to obtain CF samples.

    Techniques: SDS Page, Purification, Negative Control

    SDS-PAGE analysis of purified protein . Shown are the CaroS2K (A) and CaroS2I (B). Samples were subjected to electrophoresis in 10% polyacrylamide gels, which were stained with Coomassie blue. Lane M, molecular weight standards (kDa); lane 1, cell lysate of E. coli BL21/pET32a; lane 4, cell lysate of BL21/pET30b; lanes 2 and 5, IPTG-induced cell lysates of BL21/pES2kI and BL21/pES2I, respectively; lanes 3 and 6, purified protein obtained after elution. The arrowheads indicate the killing protein of carocin S2K (A) and the immunity protein of carocin S2I (B).

    Journal: BMC Microbiology

    Article Title: Cloning, purification, and functional characterization of Carocin S2, a ribonuclease bacteriocin produced by Pectobacterium carotovorum

    doi: 10.1186/1471-2180-11-99

    Figure Lengend Snippet: SDS-PAGE analysis of purified protein . Shown are the CaroS2K (A) and CaroS2I (B). Samples were subjected to electrophoresis in 10% polyacrylamide gels, which were stained with Coomassie blue. Lane M, molecular weight standards (kDa); lane 1, cell lysate of E. coli BL21/pET32a; lane 4, cell lysate of BL21/pET30b; lanes 2 and 5, IPTG-induced cell lysates of BL21/pES2kI and BL21/pES2I, respectively; lanes 3 and 6, purified protein obtained after elution. The arrowheads indicate the killing protein of carocin S2K (A) and the immunity protein of carocin S2I (B).

    Article Snippet: After purification through a P-100 size-exclusion column (BioRad, USA), the CaroS2K fractions were pooled and concentrated using an Amicon centriprep-50 column (Millipore, USA) and dissolved in buffer A. BL21/pES2I pellets were precipitated by ammonium sulfate (70-100%) and resuspended in buffer A. CaroS2I purification involved a similar chromatographic procedure using the Amicon centriprep-3 column (Millipore, USA).

    Techniques: SDS Page, Purification, Electrophoresis, Staining, Molecular Weight