stx2eb his  (Millipore)


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    Structured Review

    Millipore stx2eb his
    Construction of expression plasmids for Stx2e and mStx2e (A), <t>Stx2eB-His</t> (B) and Stx2eA 2 B-His (C). The schematic model shows the construction of a plasmid for the expression of target proteins. The arrow boxes indicate the open reading frames. The striped and shaded boxes indicate the start and stop codons, respectively. T7, T7 promoter; lac O, lac operator; RBS, ribosome-binding site; V5, V5 epitope tag; 6xHis, polyhistidine tag. (D) SDS-PAGE of the purified toxins and Stx2eB-derived antigens. Purified proteins (1 µ g) were resolved by 4–20% gradient SDS-PAGE and stained with Ez stain AQua. Lanes 1 and 2 show the holotoxins of Stx2e and mStx2e, respectively. The 2 bands correspond to the A 1 fragment of the A subunit (27.3 kDa) and the B subunit (7.6 kDa). Lanes 3 and 4 show Stx2eA 2 B-His and Stx2eB-His, respectively. The protein bands corresponding to the B subunit are slightly larger than the native B subunit, because of the additional 6xHis tag. The putative A 2 fragment of the A subunit (5.8 kDa) is not visible in the holotoxin or Stx2eA 2 B-His samples. Lane M shows the molecular size markers.
    Stx2eb His, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stx2eb his/product/Millipore
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    stx2eb his - by Bioz Stars, 2020-05
    85/100 stars

    Images

    1) Product Images from "Evaluation of Recombinant Forms of the Shiga Toxin Variant Stx2eB Subunit and Non-Toxic Mutant Stx2e as Vaccine Candidates against Porcine Edema Disease"

    Article Title: Evaluation of Recombinant Forms of the Shiga Toxin Variant Stx2eB Subunit and Non-Toxic Mutant Stx2e as Vaccine Candidates against Porcine Edema Disease

    Journal: The Journal of Veterinary Medical Science

    doi: 10.1292/jvms.13-0118

    Construction of expression plasmids for Stx2e and mStx2e (A), Stx2eB-His (B) and Stx2eA 2 B-His (C). The schematic model shows the construction of a plasmid for the expression of target proteins. The arrow boxes indicate the open reading frames. The striped and shaded boxes indicate the start and stop codons, respectively. T7, T7 promoter; lac O, lac operator; RBS, ribosome-binding site; V5, V5 epitope tag; 6xHis, polyhistidine tag. (D) SDS-PAGE of the purified toxins and Stx2eB-derived antigens. Purified proteins (1 µ g) were resolved by 4–20% gradient SDS-PAGE and stained with Ez stain AQua. Lanes 1 and 2 show the holotoxins of Stx2e and mStx2e, respectively. The 2 bands correspond to the A 1 fragment of the A subunit (27.3 kDa) and the B subunit (7.6 kDa). Lanes 3 and 4 show Stx2eA 2 B-His and Stx2eB-His, respectively. The protein bands corresponding to the B subunit are slightly larger than the native B subunit, because of the additional 6xHis tag. The putative A 2 fragment of the A subunit (5.8 kDa) is not visible in the holotoxin or Stx2eA 2 B-His samples. Lane M shows the molecular size markers.
    Figure Legend Snippet: Construction of expression plasmids for Stx2e and mStx2e (A), Stx2eB-His (B) and Stx2eA 2 B-His (C). The schematic model shows the construction of a plasmid for the expression of target proteins. The arrow boxes indicate the open reading frames. The striped and shaded boxes indicate the start and stop codons, respectively. T7, T7 promoter; lac O, lac operator; RBS, ribosome-binding site; V5, V5 epitope tag; 6xHis, polyhistidine tag. (D) SDS-PAGE of the purified toxins and Stx2eB-derived antigens. Purified proteins (1 µ g) were resolved by 4–20% gradient SDS-PAGE and stained with Ez stain AQua. Lanes 1 and 2 show the holotoxins of Stx2e and mStx2e, respectively. The 2 bands correspond to the A 1 fragment of the A subunit (27.3 kDa) and the B subunit (7.6 kDa). Lanes 3 and 4 show Stx2eA 2 B-His and Stx2eB-His, respectively. The protein bands corresponding to the B subunit are slightly larger than the native B subunit, because of the additional 6xHis tag. The putative A 2 fragment of the A subunit (5.8 kDa) is not visible in the holotoxin or Stx2eA 2 B-His samples. Lane M shows the molecular size markers.

    Techniques Used: Expressing, Plasmid Preparation, Binding Assay, SDS Page, Purification, Derivative Assay, Staining

    Anti-Stx2e IgG titer and lethal toxin-neutralization activity in sera from individual mice immunized i.p. with mStx2e (●), Stx2eA 2 B-His (■) and Stx2eB-His (♦). All serum samples from immunized mice on day 43 were assessed for anti-Stx2e IgG titer as well as ED 50 titer. Open symbols indicate sera that gave ED 50 titers≥2, which was the cut-off for positive Stx2e-neutralizing activity.
    Figure Legend Snippet: Anti-Stx2e IgG titer and lethal toxin-neutralization activity in sera from individual mice immunized i.p. with mStx2e (●), Stx2eA 2 B-His (■) and Stx2eB-His (♦). All serum samples from immunized mice on day 43 were assessed for anti-Stx2e IgG titer as well as ED 50 titer. Open symbols indicate sera that gave ED 50 titers≥2, which was the cut-off for positive Stx2e-neutralizing activity.

    Techniques Used: Neutralization, Activity Assay, Mouse Assay

    A. Survival profile of mice immunized i.n. with mStx2e (●), Stx2eA 2 B-His (■) or Stx2eB-His (♦), after i.p. injection with 0.1 µ g of the native toxin Stx2e. Mice administered PBS (▲) were used as negative controls. B. Anti-Stx2e IgG titers in individual sera and protection from lethal toxin challenge for mice immunized i.n. with mStx2e (●), Stx2eA 2 B-His (■) and Stx2eB-His (♦). All serum samples from immunized mice on day 178 were assessed for anti-Stx2e IgG titer. Open symbols represent sera from protected mice.
    Figure Legend Snippet: A. Survival profile of mice immunized i.n. with mStx2e (●), Stx2eA 2 B-His (■) or Stx2eB-His (♦), after i.p. injection with 0.1 µ g of the native toxin Stx2e. Mice administered PBS (▲) were used as negative controls. B. Anti-Stx2e IgG titers in individual sera and protection from lethal toxin challenge for mice immunized i.n. with mStx2e (●), Stx2eA 2 B-His (■) and Stx2eB-His (♦). All serum samples from immunized mice on day 178 were assessed for anti-Stx2e IgG titer. Open symbols represent sera from protected mice.

    Techniques Used: Mouse Assay, Injection

    Cytokines in the sera of Stx2e-challenged mice. Five mice per group were immunized with mStx2e (A), Stx2eA 2 B-His (B), Stx2eB-His (C) or PBS (D). Sera were collected before and three days after lethal challenge with Stx2e. Concentrations of IL-10, IL-12, IL-4 and IL-5 before and after Stx2e challenge are shown as log 10 -fold change ± standard deviations (SD). Levels of IL-2, IL-17, GM-CSF, IFN-γ and TNF-α were unchanged and are not shown. An asterisk denotes significance at P
    Figure Legend Snippet: Cytokines in the sera of Stx2e-challenged mice. Five mice per group were immunized with mStx2e (A), Stx2eA 2 B-His (B), Stx2eB-His (C) or PBS (D). Sera were collected before and three days after lethal challenge with Stx2e. Concentrations of IL-10, IL-12, IL-4 and IL-5 before and after Stx2e challenge are shown as log 10 -fold change ± standard deviations (SD). Levels of IL-2, IL-17, GM-CSF, IFN-γ and TNF-α were unchanged and are not shown. An asterisk denotes significance at P

    Techniques Used: Mouse Assay

    2) Product Images from "Evaluation of Recombinant Forms of the Shiga Toxin Variant Stx2eB Subunit and Non-Toxic Mutant Stx2e as Vaccine Candidates against Porcine Edema Disease"

    Article Title: Evaluation of Recombinant Forms of the Shiga Toxin Variant Stx2eB Subunit and Non-Toxic Mutant Stx2e as Vaccine Candidates against Porcine Edema Disease

    Journal: The Journal of Veterinary Medical Science

    doi: 10.1292/jvms.13-0118

    Construction of expression plasmids for Stx2e and mStx2e (A), Stx2eB-His (B) and Stx2eA 2 B-His (C). The schematic model shows the construction of a plasmid for the expression of target proteins. The arrow boxes indicate the open reading frames. The striped and shaded boxes indicate the start and stop codons, respectively. T7, T7 promoter; lac O, lac operator; RBS, ribosome-binding site; V5, V5 epitope tag; 6xHis, polyhistidine tag. (D) SDS-PAGE of the purified toxins and Stx2eB-derived antigens. Purified proteins (1 µ g) were resolved by 4–20% gradient SDS-PAGE and stained with Ez stain AQua. Lanes 1 and 2 show the holotoxins of Stx2e and mStx2e, respectively. The 2 bands correspond to the A 1 fragment of the A subunit (27.3 kDa) and the B subunit (7.6 kDa). Lanes 3 and 4 show Stx2eA 2 B-His and Stx2eB-His, respectively. The protein bands corresponding to the B subunit are slightly larger than the native B subunit, because of the additional 6xHis tag. The putative A 2 fragment of the A subunit (5.8 kDa) is not visible in the holotoxin or Stx2eA 2 B-His samples. Lane M shows the molecular size markers.
    Figure Legend Snippet: Construction of expression plasmids for Stx2e and mStx2e (A), Stx2eB-His (B) and Stx2eA 2 B-His (C). The schematic model shows the construction of a plasmid for the expression of target proteins. The arrow boxes indicate the open reading frames. The striped and shaded boxes indicate the start and stop codons, respectively. T7, T7 promoter; lac O, lac operator; RBS, ribosome-binding site; V5, V5 epitope tag; 6xHis, polyhistidine tag. (D) SDS-PAGE of the purified toxins and Stx2eB-derived antigens. Purified proteins (1 µ g) were resolved by 4–20% gradient SDS-PAGE and stained with Ez stain AQua. Lanes 1 and 2 show the holotoxins of Stx2e and mStx2e, respectively. The 2 bands correspond to the A 1 fragment of the A subunit (27.3 kDa) and the B subunit (7.6 kDa). Lanes 3 and 4 show Stx2eA 2 B-His and Stx2eB-His, respectively. The protein bands corresponding to the B subunit are slightly larger than the native B subunit, because of the additional 6xHis tag. The putative A 2 fragment of the A subunit (5.8 kDa) is not visible in the holotoxin or Stx2eA 2 B-His samples. Lane M shows the molecular size markers.

    Techniques Used: Expressing, Plasmid Preparation, Binding Assay, SDS Page, Purification, Derivative Assay, Staining

    Anti-Stx2e IgG titer and lethal toxin-neutralization activity in sera from individual mice immunized i.p. with mStx2e (●), Stx2eA 2 B-His (■) and Stx2eB-His (♦). All serum samples from immunized mice on day 43 were assessed for anti-Stx2e IgG titer as well as ED 50 titer. Open symbols indicate sera that gave ED 50 titers≥2, which was the cut-off for positive Stx2e-neutralizing activity.
    Figure Legend Snippet: Anti-Stx2e IgG titer and lethal toxin-neutralization activity in sera from individual mice immunized i.p. with mStx2e (●), Stx2eA 2 B-His (■) and Stx2eB-His (♦). All serum samples from immunized mice on day 43 were assessed for anti-Stx2e IgG titer as well as ED 50 titer. Open symbols indicate sera that gave ED 50 titers≥2, which was the cut-off for positive Stx2e-neutralizing activity.

    Techniques Used: Neutralization, Activity Assay, Mouse Assay

    A. Survival profile of mice immunized i.n. with mStx2e (●), Stx2eA 2 B-His (■) or Stx2eB-His (♦), after i.p. injection with 0.1 µ g of the native toxin Stx2e. Mice administered PBS (▲) were used as negative controls. B. Anti-Stx2e IgG titers in individual sera and protection from lethal toxin challenge for mice immunized i.n. with mStx2e (●), Stx2eA 2 B-His (■) and Stx2eB-His (♦). All serum samples from immunized mice on day 178 were assessed for anti-Stx2e IgG titer. Open symbols represent sera from protected mice.
    Figure Legend Snippet: A. Survival profile of mice immunized i.n. with mStx2e (●), Stx2eA 2 B-His (■) or Stx2eB-His (♦), after i.p. injection with 0.1 µ g of the native toxin Stx2e. Mice administered PBS (▲) were used as negative controls. B. Anti-Stx2e IgG titers in individual sera and protection from lethal toxin challenge for mice immunized i.n. with mStx2e (●), Stx2eA 2 B-His (■) and Stx2eB-His (♦). All serum samples from immunized mice on day 178 were assessed for anti-Stx2e IgG titer. Open symbols represent sera from protected mice.

    Techniques Used: Mouse Assay, Injection

    Cytokines in the sera of Stx2e-challenged mice. Five mice per group were immunized with mStx2e (A), Stx2eA 2 B-His (B), Stx2eB-His (C) or PBS (D). Sera were collected before and three days after lethal challenge with Stx2e. Concentrations of IL-10, IL-12, IL-4 and IL-5 before and after Stx2e challenge are shown as log 10 -fold change ± standard deviations (SD). Levels of IL-2, IL-17, GM-CSF, IFN-γ and TNF-α were unchanged and are not shown. An asterisk denotes significance at P
    Figure Legend Snippet: Cytokines in the sera of Stx2e-challenged mice. Five mice per group were immunized with mStx2e (A), Stx2eA 2 B-His (B), Stx2eB-His (C) or PBS (D). Sera were collected before and three days after lethal challenge with Stx2e. Concentrations of IL-10, IL-12, IL-4 and IL-5 before and after Stx2e challenge are shown as log 10 -fold change ± standard deviations (SD). Levels of IL-2, IL-17, GM-CSF, IFN-γ and TNF-α were unchanged and are not shown. An asterisk denotes significance at P

    Techniques Used: Mouse Assay

    Related Articles

    Transformation Assay:

    Article Title: Evaluation of Recombinant Forms of the Shiga Toxin Variant Stx2eB Subunit and Non-Toxic Mutant Stx2e as Vaccine Candidates against Porcine Edema Disease
    Article Snippet: .. Stx2eB-His and Stx2eA2 B-His protein production andpurification : Production of Stx2eB-His was performed as described for Stx2e and mStx2e, except E. coli NovaBlue (Novagen, Merck KGaA, Darmstadt, Germany) were transformed with the plasmid pETStx2eB-His. .. For production of Stx2eA2 B,E. coli BL21 Star (DE3) were transformed with pETStx2eA2 B-His and cultured in Plusgrow broth (Nacalai Tesque, Kyoto, Japan) in the presence of ampicillin (50 µ g/ml ).

    Plasmid Preparation:

    Article Title: Evaluation of Recombinant Forms of the Shiga Toxin Variant Stx2eB Subunit and Non-Toxic Mutant Stx2e as Vaccine Candidates against Porcine Edema Disease
    Article Snippet: .. Stx2eB-His and Stx2eA2 B-His protein production andpurification : Production of Stx2eB-His was performed as described for Stx2e and mStx2e, except E. coli NovaBlue (Novagen, Merck KGaA, Darmstadt, Germany) were transformed with the plasmid pETStx2eB-His. .. For production of Stx2eA2 B,E. coli BL21 Star (DE3) were transformed with pETStx2eA2 B-His and cultured in Plusgrow broth (Nacalai Tesque, Kyoto, Japan) in the presence of ampicillin (50 µ g/ml ).