e8200  (New England Biolabs)


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    Name:
    pMAL Protein Fusion and Purification System
    Description:
    pMAL Protein Fusion and Purification System
    Catalog Number:
    E8200S
    Price:
    677
    Category:
    E coli Protein Expression Kits
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    Structured Review

    New England Biolabs e8200
    pMAL Protein Fusion and Purification System
    pMAL Protein Fusion and Purification System
    https://www.bioz.com/result/e8200/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    e8200 - by Bioz Stars, 2021-06
    99/100 stars

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    Related Articles

    Over Expression:

    Article Title: Molecular and functional insights into a novel teleost malectin from big-belly seahorse Hippocampus abdominalis
    Article Snippet: .. 3.4 Overexpression and purification of rHaMLEC rHaMLEC was overexpressed in E. coli BL21(DE3) cells and purified using a pMAL protein fusion and purification system (New England Biolabs, USA). .. The following samples were analyzed by SDS-PAGE throughout the purification steps: total protein extract after sonication, supernatant after centrifugation, and purified protein ( ).

    Purification:

    Article Title: Molecular and functional insights into a novel teleost malectin from big-belly seahorse Hippocampus abdominalis
    Article Snippet: .. 3.4 Overexpression and purification of rHaMLEC rHaMLEC was overexpressed in E. coli BL21(DE3) cells and purified using a pMAL protein fusion and purification system (New England Biolabs, USA). .. The following samples were analyzed by SDS-PAGE throughout the purification steps: total protein extract after sonication, supernatant after centrifugation, and purified protein ( ).

    Article Title: A valid strategy for precise identifications of transcription factor binding sites in combinatorial regulation using bioinformatic and experimental approaches
    Article Snippet: The luciferase activities were detected by a Glomax 20/20 luminometer following the manufactory’s instruction on Dual-Luciferase® Reporter (DLR™) Assay System (Promega). .. Protein expression and purification Constructs were built from pMAL-c2G (Additional file ), as instructed for the pMAL™ protein fusion and purification system (New England Biolabs). .. The host strain E . coli BL21 (DE3) was transformed by the target vector and grown at 37 C in LB solution with ampicillin (50 μg/ml) and chloramphenicol (50 μg/ml), and added with IPTG to 0.5 mM when the solution reached 0.5 at OD600, then cultured under 25 C for six hours before protein extraction.

    Article Title: The Integrative Conjugative Element (ICE) of Mycoplasma agalactiae: Key Elements Involved in Horizontal Dissemination and Influence of Coresident ICEs
    Article Snippet: .. The anti-CDS14 lipoprotein rabbit serum was produced by animal immunization with a recombinant CDS14 protein (pMAL protein fusion and purification system; New England Biolabs). ..

    Article Title: The PtlE Protein of Bordetella pertussis Has Peptidoglycanase Activity Required for Ptl-Mediated Pertussis Toxin Secretion
    Article Snippet: The PCR product was ligated into plasmid pMAL-p2x by using the Xba I and Hin dIII sites. .. A maltose-binding protein fusion to this region of PtlF was generated and purified by using the pMAL protein fusion and purification system (New England BioLabs, Inc.). .. The fusion protein was expressed in E. coli UT5600 to increase expression levels and decrease proteolytic degradation.

    Article Title: Nonimmune Binding of Human Immunoglobulin A (IgA) and IgG Fc by Distinct Sequence Segments of the EibF Cell Surface Protein of Escherichia coli
    Article Snippet: .. Our choice was the pMAL-c2X protein fusion and purification system (New England Biolabs). .. The strategy involved fusing an eib gene to the N terminus of the malE gene of the plasmid vector pMal-c2X.

    Expressing:

    Article Title: A valid strategy for precise identifications of transcription factor binding sites in combinatorial regulation using bioinformatic and experimental approaches
    Article Snippet: The luciferase activities were detected by a Glomax 20/20 luminometer following the manufactory’s instruction on Dual-Luciferase® Reporter (DLR™) Assay System (Promega). .. Protein expression and purification Constructs were built from pMAL-c2G (Additional file ), as instructed for the pMAL™ protein fusion and purification system (New England Biolabs). .. The host strain E . coli BL21 (DE3) was transformed by the target vector and grown at 37 C in LB solution with ampicillin (50 μg/ml) and chloramphenicol (50 μg/ml), and added with IPTG to 0.5 mM when the solution reached 0.5 at OD600, then cultured under 25 C for six hours before protein extraction.

    Construct:

    Article Title: A valid strategy for precise identifications of transcription factor binding sites in combinatorial regulation using bioinformatic and experimental approaches
    Article Snippet: The luciferase activities were detected by a Glomax 20/20 luminometer following the manufactory’s instruction on Dual-Luciferase® Reporter (DLR™) Assay System (Promega). .. Protein expression and purification Constructs were built from pMAL-c2G (Additional file ), as instructed for the pMAL™ protein fusion and purification system (New England Biolabs). .. The host strain E . coli BL21 (DE3) was transformed by the target vector and grown at 37 C in LB solution with ampicillin (50 μg/ml) and chloramphenicol (50 μg/ml), and added with IPTG to 0.5 mM when the solution reached 0.5 at OD600, then cultured under 25 C for six hours before protein extraction.

    Produced:

    Article Title: The Integrative Conjugative Element (ICE) of Mycoplasma agalactiae: Key Elements Involved in Horizontal Dissemination and Influence of Coresident ICEs
    Article Snippet: .. The anti-CDS14 lipoprotein rabbit serum was produced by animal immunization with a recombinant CDS14 protein (pMAL protein fusion and purification system; New England Biolabs). ..

    Recombinant:

    Article Title: The Integrative Conjugative Element (ICE) of Mycoplasma agalactiae: Key Elements Involved in Horizontal Dissemination and Influence of Coresident ICEs
    Article Snippet: .. The anti-CDS14 lipoprotein rabbit serum was produced by animal immunization with a recombinant CDS14 protein (pMAL protein fusion and purification system; New England Biolabs). ..

    Sonication:

    Article Title: Mutants of the Zinc Ligands of Lacticin 481 Synthetase Retain Dehydration Activity but Have Impaired Cyclization Activity †
    Article Snippet: .. Cells were lysed by sonication on ice and then centrifuged at 23,700 x g for 30 min at 4 o C. The supernatant was filtered through a 0.45 μm cellulose acetate filter onto a chitin (NEB, 20 mL bed volume) column equilibrated with 100 mM HEPES, pH 7.2, 500 mM NaCl and 1 mM EDTA at 4 o C. The fusion protein was allowed to bind to the chitin column for 2 h with gentle shaking. ..

    Generated:

    Article Title: The PtlE Protein of Bordetella pertussis Has Peptidoglycanase Activity Required for Ptl-Mediated Pertussis Toxin Secretion
    Article Snippet: The PCR product was ligated into plasmid pMAL-p2x by using the Xba I and Hin dIII sites. .. A maltose-binding protein fusion to this region of PtlF was generated and purified by using the pMAL protein fusion and purification system (New England BioLabs, Inc.). .. The fusion protein was expressed in E. coli UT5600 to increase expression levels and decrease proteolytic degradation.

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    New England Biolabs pmal c2x protein fusion
    MBP-EibA fusions. Broth cultures were grown and induced with IPTG, and whole-cell were extracts prepared as described in Materials and Methods. Equivalent amounts of cell extracts were fractionated by SDS-PAGE (8% acrylamide) and blotted to PVDF. The blot was incubated first with human IgG Fc-HRP (A) and second with rabbit anti-MBP developed with donkey anti-rabbit Ig (B) (see Materials and Methods). Lanes: 1, <t>pMal-c2X;</t> 2, EibA (amino acids [aa] 254 to 392), pDC2240; 3, EibA (aa 254 to 344), pDC2283; 4, EibA (aa 300 to 392), pDC2252; 5, EibA (aa 254 to 298), pCS7299; 6, EibF (aa 318 to 459), pCS7280.
    Pmal C2x Protein Fusion, 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/pmal c2x protein fusion/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pmal c2x protein fusion - by Bioz Stars, 2021-06
    99/100 stars
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    MBP-EibA fusions. Broth cultures were grown and induced with IPTG, and whole-cell were extracts prepared as described in Materials and Methods. Equivalent amounts of cell extracts were fractionated by SDS-PAGE (8% acrylamide) and blotted to PVDF. The blot was incubated first with human IgG Fc-HRP (A) and second with rabbit anti-MBP developed with donkey anti-rabbit Ig (B) (see Materials and Methods). Lanes: 1, pMal-c2X; 2, EibA (amino acids [aa] 254 to 392), pDC2240; 3, EibA (aa 254 to 344), pDC2283; 4, EibA (aa 300 to 392), pDC2252; 5, EibA (aa 254 to 298), pCS7299; 6, EibF (aa 318 to 459), pCS7280.

    Journal: Infection and Immunity

    Article Title: Nonimmune Binding of Human Immunoglobulin A (IgA) and IgG Fc by Distinct Sequence Segments of the EibF Cell Surface Protein of Escherichia coli

    doi: 10.1128/IAI.69.12.7293-7303.2001

    Figure Lengend Snippet: MBP-EibA fusions. Broth cultures were grown and induced with IPTG, and whole-cell were extracts prepared as described in Materials and Methods. Equivalent amounts of cell extracts were fractionated by SDS-PAGE (8% acrylamide) and blotted to PVDF. The blot was incubated first with human IgG Fc-HRP (A) and second with rabbit anti-MBP developed with donkey anti-rabbit Ig (B) (see Materials and Methods). Lanes: 1, pMal-c2X; 2, EibA (amino acids [aa] 254 to 392), pDC2240; 3, EibA (aa 254 to 344), pDC2283; 4, EibA (aa 300 to 392), pDC2252; 5, EibA (aa 254 to 298), pCS7299; 6, EibF (aa 318 to 459), pCS7280.

    Article Snippet: Our choice was the pMAL-c2X protein fusion and purification system (New England Biolabs).

    Techniques: SDS Page, Incubation

    Protein expression of PG2-ICEA mutants. (A) Immunostaining of M. agalactiae colonies showing CDS14 lipoprotein expression at the surface of 5632 cells. Colony blotting was carried out by using a specific serum (anti-CDS14), and ICEA-negative PG2 cells (PG2) were used as a negative control. (B) Western blot analysis of CDS14 lipoprotein expression in 5632 and PG2 ICEA cells. CDS14 lipoprotein expression in strain 5632 containing three chromosomal ICEA copies (lane 1) was not abrogated in a 5632 mutant harboring a cds14 knockout ICEA copy (lane 2). CDS14 lipoprotein expression was detectable in PG2 transconjugants that had acquired a mutant ICEA harboring an mTn inserted in ncr19 / E (lane 4) but not in strain PG2 (lane 3) or in PG2 transconjugants harboring a cds14 knockout ICEA (lane 5). Transformation of PG2 transconjugants harboring a cds14 knockout ICEA with a plasmid expressing CDS14 restored the expression of the lipoprotein (lane 6). A specific serum raised against lipoprotein P80 was used as a control (P80). (C) Schematic illustrating the protein expression profiles of selected mutant ICEAs in PG2 cells. Mutant ICEAs are identified according to Table S2 , and ICEA products detected by proteomics ( Table S4 ) are indicated (closed arrows).

    Journal: mBio

    Article Title: The Integrative Conjugative Element (ICE) of Mycoplasma agalactiae: Key Elements Involved in Horizontal Dissemination and Influence of Coresident ICEs

    doi: 10.1128/mBio.00873-18

    Figure Lengend Snippet: Protein expression of PG2-ICEA mutants. (A) Immunostaining of M. agalactiae colonies showing CDS14 lipoprotein expression at the surface of 5632 cells. Colony blotting was carried out by using a specific serum (anti-CDS14), and ICEA-negative PG2 cells (PG2) were used as a negative control. (B) Western blot analysis of CDS14 lipoprotein expression in 5632 and PG2 ICEA cells. CDS14 lipoprotein expression in strain 5632 containing three chromosomal ICEA copies (lane 1) was not abrogated in a 5632 mutant harboring a cds14 knockout ICEA copy (lane 2). CDS14 lipoprotein expression was detectable in PG2 transconjugants that had acquired a mutant ICEA harboring an mTn inserted in ncr19 / E (lane 4) but not in strain PG2 (lane 3) or in PG2 transconjugants harboring a cds14 knockout ICEA (lane 5). Transformation of PG2 transconjugants harboring a cds14 knockout ICEA with a plasmid expressing CDS14 restored the expression of the lipoprotein (lane 6). A specific serum raised against lipoprotein P80 was used as a control (P80). (C) Schematic illustrating the protein expression profiles of selected mutant ICEAs in PG2 cells. Mutant ICEAs are identified according to Table S2 , and ICEA products detected by proteomics ( Table S4 ) are indicated (closed arrows).

    Article Snippet: The anti-CDS14 lipoprotein rabbit serum was produced by animal immunization with a recombinant CDS14 protein (pMAL protein fusion and purification system; New England Biolabs).

    Techniques: Expressing, Immunostaining, Negative Control, Western Blot, Mutagenesis, Knock-Out, Transformation Assay, Plasmid Preparation

    Overview of conjugative ICE transfer in M. agalactiae . This schematic illustrates the 5 key steps in ICEA transfer based on current knowledge in other bacteria ( 1 , 18 ). Under normal conditions, ICEA copies are found integrated into the host chromosome and most ICEA genes are not expressed. Among the few proteins expressed by chromosomal ICEAs is the CDS14 lipoprotein, which is surface exposed and plays a critical role in initiating the conjugative process (step 1). When ICEA gene expression is induced, under specific cellular conditions or stochastically, the cis -acting DDE transposase is produced and one of the three ICEA copies excises from the chromosome and forms a circular double-stranded DNA (dsDNA) molecule (step 2). ICEA circularization induces the expression of the conjugative module, whose products assemble into the mating pore, a simplified form of type IV secretion system (T4SS) found in more-complex bacteria (step 3). A protein complex known as a relaxosome recognizes the origin of transfer ( oriT ) on the circular ICEA, and a relaxase generates a linear single-stranded DNA (ssDNA) by nicking the ICEA DNA (step 4). Finally, the relaxosome complex interacts with the TraG-like (VirD4 homologue) energetic component found at the inner side of the membrane that facilitates the transfer of the ssDNA bound to the relaxase through the mating channel (step 5). Once in the recipient strain, the ICEA recircularizes, becomes doubly stranded, and integrates randomly into the host chromosome. The minimal functional ICEA encompasses 80% of the coding sequence and includes a gene cluster ( cds5 to cds19 , encoding proteins with transmembrane domains) that most likely represents a module associated with the conjugative channel. Additional essential ICEA determinants included the CDS14 surface lipoprotein, the CDSG putative partitioning protein, and the DDE transposase (CDS22), together with several proteins of unknown function (CDS1, CDSA, CDSC, and CDS30).

    Journal: mBio

    Article Title: The Integrative Conjugative Element (ICE) of Mycoplasma agalactiae: Key Elements Involved in Horizontal Dissemination and Influence of Coresident ICEs

    doi: 10.1128/mBio.00873-18

    Figure Lengend Snippet: Overview of conjugative ICE transfer in M. agalactiae . This schematic illustrates the 5 key steps in ICEA transfer based on current knowledge in other bacteria ( 1 , 18 ). Under normal conditions, ICEA copies are found integrated into the host chromosome and most ICEA genes are not expressed. Among the few proteins expressed by chromosomal ICEAs is the CDS14 lipoprotein, which is surface exposed and plays a critical role in initiating the conjugative process (step 1). When ICEA gene expression is induced, under specific cellular conditions or stochastically, the cis -acting DDE transposase is produced and one of the three ICEA copies excises from the chromosome and forms a circular double-stranded DNA (dsDNA) molecule (step 2). ICEA circularization induces the expression of the conjugative module, whose products assemble into the mating pore, a simplified form of type IV secretion system (T4SS) found in more-complex bacteria (step 3). A protein complex known as a relaxosome recognizes the origin of transfer ( oriT ) on the circular ICEA, and a relaxase generates a linear single-stranded DNA (ssDNA) by nicking the ICEA DNA (step 4). Finally, the relaxosome complex interacts with the TraG-like (VirD4 homologue) energetic component found at the inner side of the membrane that facilitates the transfer of the ssDNA bound to the relaxase through the mating channel (step 5). Once in the recipient strain, the ICEA recircularizes, becomes doubly stranded, and integrates randomly into the host chromosome. The minimal functional ICEA encompasses 80% of the coding sequence and includes a gene cluster ( cds5 to cds19 , encoding proteins with transmembrane domains) that most likely represents a module associated with the conjugative channel. Additional essential ICEA determinants included the CDS14 surface lipoprotein, the CDSG putative partitioning protein, and the DDE transposase (CDS22), together with several proteins of unknown function (CDS1, CDSA, CDSC, and CDS30).

    Article Snippet: The anti-CDS14 lipoprotein rabbit serum was produced by animal immunization with a recombinant CDS14 protein (pMAL protein fusion and purification system; New England Biolabs).

    Techniques: Expressing, Produced, Functional Assay, Sequencing

    SDS-PAGE validation of purified rHaMLEC fusion protein. Lane 1 = protein size marker (Enzynomics-Korea); Lane 2 = supernatant from E. coli BL21 (DE3) transfected with rHaMLEC-pMAL-c5X; Lane 3 = total extract from IPTG-induced E. coli BL21 (DE3) cells transfected with rHaMLEC-pMAL-c5X; Lane 4 = supernatant of IPTG-induced E. coli BL21 (DE3) cells transfected with rHaMLEC-pMAL-c5X; Lane 5 = purified rHaMLEC-MBP fusion protein.

    Journal: Fish & Shellfish Immunology

    Article Title: Molecular and functional insights into a novel teleost malectin from big-belly seahorse Hippocampus abdominalis

    doi: 10.1016/j.fsi.2020.02.044

    Figure Lengend Snippet: SDS-PAGE validation of purified rHaMLEC fusion protein. Lane 1 = protein size marker (Enzynomics-Korea); Lane 2 = supernatant from E. coli BL21 (DE3) transfected with rHaMLEC-pMAL-c5X; Lane 3 = total extract from IPTG-induced E. coli BL21 (DE3) cells transfected with rHaMLEC-pMAL-c5X; Lane 4 = supernatant of IPTG-induced E. coli BL21 (DE3) cells transfected with rHaMLEC-pMAL-c5X; Lane 5 = purified rHaMLEC-MBP fusion protein.

    Article Snippet: 3.4 Overexpression and purification of rHaMLEC rHaMLEC was overexpressed in E. coli BL21(DE3) cells and purified using a pMAL protein fusion and purification system (New England Biolabs, USA).

    Techniques: SDS Page, Purification, Marker, Transfection

    Overview of conjugative ICE transfer in M. agalactiae ). Under normal conditions, ICEA copies are found integrated into the host chromosome and most ICEA genes are not expressed. Among the few proteins expressed by chromosomal ICEAs is the CDS14 lipoprotein, which is surface exposed and plays a critical role in initiating the conjugative process (step 1). When ICEA gene expression is induced, under specific cellular conditions or stochastically, the cis -acting DDE transposase is produced and one of the three ICEA copies excises from the chromosome and forms a circular double-stranded DNA (dsDNA) molecule (step 2). ICEA circularization induces the expression of the conjugative module, whose products assemble into the mating pore, a simplified form of type IV secretion system (T4SS) found in more-complex bacteria (step 3). A protein complex known as a relaxosome recognizes the origin of transfer ( oriT ) on the circular ICEA, and a relaxase generates a linear single-stranded DNA (ssDNA) by nicking the ICEA DNA (step 4). Finally, the relaxosome complex interacts with the TraG-like (VirD4 homologue) energetic component found at the inner side of the membrane that facilitates the transfer of the ssDNA bound to the relaxase through the mating channel (step 5). Once in the recipient strain, the ICEA recircularizes, becomes doubly stranded, and integrates randomly into the host chromosome. The minimal functional ICEA encompasses 80% of the coding sequence and includes a gene cluster ( cds5 to cds19 , encoding proteins with transmembrane domains) that most likely represents a module associated with the conjugative channel. Additional essential ICEA determinants included the CDS14 surface lipoprotein, the CDSG putative partitioning protein, and the DDE transposase (CDS22), together with several proteins of unknown function (CDS1, CDSA, CDSC, and CDS30).

    Journal: mBio

    Article Title: The Integrative Conjugative Element (ICE) of Mycoplasma agalactiae: Key Elements Involved in Horizontal Dissemination and Influence of Coresident ICEs

    doi: 10.1128/mBio.00873-18

    Figure Lengend Snippet: Overview of conjugative ICE transfer in M. agalactiae ). Under normal conditions, ICEA copies are found integrated into the host chromosome and most ICEA genes are not expressed. Among the few proteins expressed by chromosomal ICEAs is the CDS14 lipoprotein, which is surface exposed and plays a critical role in initiating the conjugative process (step 1). When ICEA gene expression is induced, under specific cellular conditions or stochastically, the cis -acting DDE transposase is produced and one of the three ICEA copies excises from the chromosome and forms a circular double-stranded DNA (dsDNA) molecule (step 2). ICEA circularization induces the expression of the conjugative module, whose products assemble into the mating pore, a simplified form of type IV secretion system (T4SS) found in more-complex bacteria (step 3). A protein complex known as a relaxosome recognizes the origin of transfer ( oriT ) on the circular ICEA, and a relaxase generates a linear single-stranded DNA (ssDNA) by nicking the ICEA DNA (step 4). Finally, the relaxosome complex interacts with the TraG-like (VirD4 homologue) energetic component found at the inner side of the membrane that facilitates the transfer of the ssDNA bound to the relaxase through the mating channel (step 5). Once in the recipient strain, the ICEA recircularizes, becomes doubly stranded, and integrates randomly into the host chromosome. The minimal functional ICEA encompasses 80% of the coding sequence and includes a gene cluster ( cds5 to cds19 , encoding proteins with transmembrane domains) that most likely represents a module associated with the conjugative channel. Additional essential ICEA determinants included the CDS14 surface lipoprotein, the CDSG putative partitioning protein, and the DDE transposase (CDS22), together with several proteins of unknown function (CDS1, CDSA, CDSC, and CDS30).

    Article Snippet: The anti-CDS14 lipoprotein rabbit serum was produced by animal immunization with a recombinant CDS14 protein (pMAL protein fusion and purification system; New England Biolabs).

    Techniques: Expressing, Produced, Functional Assay, Sequencing