n. meningitidis Search Results


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  • 93
    ATCC n meningitidis
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 99 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore n meningitidis
    Ectopic reporter gene study of the neisserial thiC RS. (A) The promoter region (−35 and −10 elements) and the 5´UTR are conserved within the different Neisseria species. The transcriptional start site (TSS) is indicated with a blue arrow and the translational start codon (ATG) underlined in blue. (B) A schematic figure of the neisserial TPP RS-EGFP fusion plasmid. The genomic region corresponding to the promoter, 5´UTR and ATG start codon of thiC from N. <t>meningitidis</t> serogroups B and A, N. gonorrhoeae and N. lactamica were cloned in a translational fusion with EGFP. (C) The expression of EGFP in E. coli was analysed by Western blot, in the absence and increasing concentration of thiamine. In ΔRS background (deletion of RS region), the expression of EGFP was constitutively high regardless of additional thiamine, while the other neisserial RS clones showed thiamine-dependent EGFP expression. Empty plasmid (pEGFP), harbouring no thiC promoter and its 5´UTR was used as control. EGFP signals were quantified and normalized to the corresponding RecA signals and each set by the value of the 0 μM thiamine sample.
    N Meningitidis, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck & Co n meningitidis
    Ectopic reporter gene study of the neisserial thiC RS. (A) The promoter region (−35 and −10 elements) and the 5´UTR are conserved within the different Neisseria species. The transcriptional start site (TSS) is indicated with a blue arrow and the translational start codon (ATG) underlined in blue. (B) A schematic figure of the neisserial TPP RS-EGFP fusion plasmid. The genomic region corresponding to the promoter, 5´UTR and ATG start codon of thiC from N. <t>meningitidis</t> serogroups B and A, N. gonorrhoeae and N. lactamica were cloned in a translational fusion with EGFP. (C) The expression of EGFP in E. coli was analysed by Western blot, in the absence and increasing concentration of thiamine. In ΔRS background (deletion of RS region), the expression of EGFP was constitutively high regardless of additional thiamine, while the other neisserial RS clones showed thiamine-dependent EGFP expression. Empty plasmid (pEGFP), harbouring no thiC promoter and its 5´UTR was used as control. EGFP signals were quantified and normalized to the corresponding RecA signals and each set by the value of the 0 μM thiamine sample.
    N Meningitidis, supplied by Merck & Co, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Connaught Labs n meningitidis
    Ectopic reporter gene study of the neisserial thiC RS. (A) The promoter region (−35 and −10 elements) and the 5´UTR are conserved within the different Neisseria species. The transcriptional start site (TSS) is indicated with a blue arrow and the translational start codon (ATG) underlined in blue. (B) A schematic figure of the neisserial TPP RS-EGFP fusion plasmid. The genomic region corresponding to the promoter, 5´UTR and ATG start codon of thiC from N. <t>meningitidis</t> serogroups B and A, N. gonorrhoeae and N. lactamica were cloned in a translational fusion with EGFP. (C) The expression of EGFP in E. coli was analysed by Western blot, in the absence and increasing concentration of thiamine. In ΔRS background (deletion of RS region), the expression of EGFP was constitutively high regardless of additional thiamine, while the other neisserial RS clones showed thiamine-dependent EGFP expression. Empty plasmid (pEGFP), harbouring no thiC promoter and its 5´UTR was used as control. EGFP signals were quantified and normalized to the corresponding RecA signals and each set by the value of the 0 μM thiamine sample.
    N Meningitidis, supplied by Connaught Labs, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Virostat c n meningitidis
    Ectopic reporter gene study of the neisserial thiC RS. (A) The promoter region (−35 and −10 elements) and the 5´UTR are conserved within the different Neisseria species. The transcriptional start site (TSS) is indicated with a blue arrow and the translational start codon (ATG) underlined in blue. (B) A schematic figure of the neisserial TPP RS-EGFP fusion plasmid. The genomic region corresponding to the promoter, 5´UTR and ATG start codon of thiC from N. <t>meningitidis</t> serogroups B and A, N. gonorrhoeae and N. lactamica were cloned in a translational fusion with EGFP. (C) The expression of EGFP in E. coli was analysed by Western blot, in the absence and increasing concentration of thiamine. In ΔRS background (deletion of RS region), the expression of EGFP was constitutively high regardless of additional thiamine, while the other neisserial RS clones showed thiamine-dependent EGFP expression. Empty plasmid (pEGFP), harbouring no thiC promoter and its 5´UTR was used as control. EGFP signals were quantified and normalized to the corresponding RecA signals and each set by the value of the 0 μM thiamine sample.
    C N Meningitidis, supplied by Virostat, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson n meningitidis
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Addgene inc n meningitidis
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Copan Diagnostics n meningitidis
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis, supplied by Copan Diagnostics, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Starplex Scientific n meningitidis
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis, supplied by Starplex Scientific, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Clinical and Laboratory Standards Institute n meningitidis
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis, supplied by Clinical and Laboratory Standards Institute, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Mimotopes n meningitidis
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis, supplied by Mimotopes, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher n meningitidis
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 131 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore n meningitidis group b
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis Group B, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Santa Cruz Biotechnology n meningitidis pib
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis Pib, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco n meningitidis antiserum
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis Antiserum, supplied by Difco, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson unencapsulated n meningitidis
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    Unencapsulated N Meningitidis, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Connaught Labs n meningitidis quadrivalent
    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. <t>meningitidis</t> with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.
    N Meningitidis Quadrivalent, supplied by Connaught Labs, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Abcam wc n meningitidis
    Immunofluorescence microscopy of WCs of strains MC58, MC58Lnt and MC58LntC. To confirm the presence of meningococcal cells, cells were incubated with FITC‐labelled rabbit polyclonal IgG raised against WC N. <t>meningitidis</t> (left and right panel). To compare FHbp cell surface expression between the strains, cells were also incubated with anti‐FHbp antibody JAR4 that was detected by Alexa Fluor 555 labelled donkey anti‐mouse IgG secondary antibody (right panel).
    Wc N Meningitidis, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    US Biological Life Sciences rabbit antibody against n meningitidis
    Dynamic meningococcal disease progression selects for Opa expression. (A) Mice with mild-disease pattern were imaged over 4 days using a detection scale of 200/2000 counts. Bioluminescent signals from the mice were undetectable at 2 days (2d) post-infection. From day 3 (3d), the signals reappeared either in the brain (upper panel) or in the whole body (lower panel). Images are representative of each subgroup. Each panel follows one mouse over time. Bacterial loads in blood and CSF at indicated time points post-challenge are shown. (B) N. <t>meningitidis</t> was isolated from blood, CSF or nasal washes (N wash) at day 1, 3 and 4 after infection. Recovered isolates together with the initial inoculum, were analyzed for expression of Opa proteins (30 kDa), PilC (110 kDa), and pili (PilE subunit 16 kDa) by immunoblots. LOS (6.5 kDa) was analyzed by Tricine SDS-PAGE.
    Rabbit Antibody Against N Meningitidis, supplied by US Biological Life Sciences, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher wellcogen n meningitidis acyw135 latex reagent
    Dynamic meningococcal disease progression selects for Opa expression. (A) Mice with mild-disease pattern were imaged over 4 days using a detection scale of 200/2000 counts. Bioluminescent signals from the mice were undetectable at 2 days (2d) post-infection. From day 3 (3d), the signals reappeared either in the brain (upper panel) or in the whole body (lower panel). Images are representative of each subgroup. Each panel follows one mouse over time. Bacterial loads in blood and CSF at indicated time points post-challenge are shown. (B) N. <t>meningitidis</t> was isolated from blood, CSF or nasal washes (N wash) at day 1, 3 and 4 after infection. Recovered isolates together with the initial inoculum, were analyzed for expression of Opa proteins (30 kDa), PilC (110 kDa), and pili (PilE subunit 16 kDa) by immunoblots. LOS (6.5 kDa) was analyzed by Tricine SDS-PAGE.
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    92
    ATCC n meningitidis atcc 13090
    Dynamic meningococcal disease progression selects for Opa expression. (A) Mice with mild-disease pattern were imaged over 4 days using a detection scale of 200/2000 counts. Bioluminescent signals from the mice were undetectable at 2 days (2d) post-infection. From day 3 (3d), the signals reappeared either in the brain (upper panel) or in the whole body (lower panel). Images are representative of each subgroup. Each panel follows one mouse over time. Bacterial loads in blood and CSF at indicated time points post-challenge are shown. (B) N. <t>meningitidis</t> was isolated from blood, CSF or nasal washes (N wash) at day 1, 3 and 4 after infection. Recovered isolates together with the initial inoculum, were analyzed for expression of Opa proteins (30 kDa), PilC (110 kDa), and pili (PilE subunit 16 kDa) by immunoblots. LOS (6.5 kDa) was analyzed by Tricine SDS-PAGE.
    N Meningitidis Atcc 13090, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Promega n meningitidis strain 608b
    Bactericidal activity of protein A–purified mAb Me-1 against N. <t>meningitidis</t> strain <t>608B.</t> Guinea pig serum ( closed circles ) or heat inactivated guinea pig serum ( open circles ) were used as the source of complement to evaluate the bactericidal activity.
    N Meningitidis Strain 608b, supplied by Promega, used in various techniques. Bioz Stars score: 80/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Merck & Co n meningitidis strain c11
    Bactericidal activity of protein A–purified mAb Me-1 against N. <t>meningitidis</t> strain <t>608B.</t> Guinea pig serum ( closed circles ) or heat inactivated guinea pig serum ( open circles ) were used as the source of complement to evaluate the bactericidal activity.
    N Meningitidis Strain C11, supplied by Merck & Co, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Pasteur Institute n meningitidis strain 8013
    Bactericidal activity of protein A–purified mAb Me-1 against N. <t>meningitidis</t> strain <t>608B.</t> Guinea pig serum ( closed circles ) or heat inactivated guinea pig serum ( open circles ) were used as the source of complement to evaluate the bactericidal activity.
    N Meningitidis Strain 8013, supplied by Pasteur Institute, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Trinity Biotech polyclonal antibody against n meningitidis
    Bactericidal activity of protein A–purified mAb Me-1 against N. <t>meningitidis</t> strain <t>608B.</t> Guinea pig serum ( closed circles ) or heat inactivated guinea pig serum ( open circles ) were used as the source of complement to evaluate the bactericidal activity.
    Polyclonal Antibody Against N Meningitidis, supplied by Trinity Biotech, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. meningitidis with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.

    Journal: Biosensors & bioelectronics

    Article Title: Fully 3D Printed Integrated Reactor Array for Point-of-Care Molecular Diagnostics

    doi: 10.1016/j.bios.2018.03.009

    Figure Lengend Snippet: Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. meningitidis with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.

    Article Snippet: Bovine serum albumin (BSA), Poly (ethylene glycol) 8000 (PEG), poly(vinyl alcohol) (PVA) are from Sigma-Aldrich, RT-PCR grade water from Ambion, Inc., intercalating Eva Green fluorescent dye from Biotium, Isothermal Master Mix used in LAMP reaction buffer from ISO-001nd, OptiGene, Horsham, UK, WarmStart® colorimetric LAMP 2X master mix from New England Biolabs Inc. N. meningitidis (ATCC 13098) from American Type Culture Collection (ATCC, Rockville, MD).

    Techniques: Fluorescence, Lamp Assay, Chromatin Immunoprecipitation, Amplification, Concentration Assay

    Ectopic reporter gene study of the neisserial thiC RS. (A) The promoter region (−35 and −10 elements) and the 5´UTR are conserved within the different Neisseria species. The transcriptional start site (TSS) is indicated with a blue arrow and the translational start codon (ATG) underlined in blue. (B) A schematic figure of the neisserial TPP RS-EGFP fusion plasmid. The genomic region corresponding to the promoter, 5´UTR and ATG start codon of thiC from N. meningitidis serogroups B and A, N. gonorrhoeae and N. lactamica were cloned in a translational fusion with EGFP. (C) The expression of EGFP in E. coli was analysed by Western blot, in the absence and increasing concentration of thiamine. In ΔRS background (deletion of RS region), the expression of EGFP was constitutively high regardless of additional thiamine, while the other neisserial RS clones showed thiamine-dependent EGFP expression. Empty plasmid (pEGFP), harbouring no thiC promoter and its 5´UTR was used as control. EGFP signals were quantified and normalized to the corresponding RecA signals and each set by the value of the 0 μM thiamine sample.

    Journal: RNA Biology

    Article Title: Characterization of a transcriptional TPP riboswitch in the human pathogen Neisseria meningitidis

    doi: 10.1080/15476286.2020.1727188

    Figure Lengend Snippet: Ectopic reporter gene study of the neisserial thiC RS. (A) The promoter region (−35 and −10 elements) and the 5´UTR are conserved within the different Neisseria species. The transcriptional start site (TSS) is indicated with a blue arrow and the translational start codon (ATG) underlined in blue. (B) A schematic figure of the neisserial TPP RS-EGFP fusion plasmid. The genomic region corresponding to the promoter, 5´UTR and ATG start codon of thiC from N. meningitidis serogroups B and A, N. gonorrhoeae and N. lactamica were cloned in a translational fusion with EGFP. (C) The expression of EGFP in E. coli was analysed by Western blot, in the absence and increasing concentration of thiamine. In ΔRS background (deletion of RS region), the expression of EGFP was constitutively high regardless of additional thiamine, while the other neisserial RS clones showed thiamine-dependent EGFP expression. Empty plasmid (pEGFP), harbouring no thiC promoter and its 5´UTR was used as control. EGFP signals were quantified and normalized to the corresponding RecA signals and each set by the value of the 0 μM thiamine sample.

    Article Snippet: In order to assess the effect of different thiamine sources on the TPP-mediated expression of thiC in E. coli reporter strains and N. meningitidis , the following experimental procedures were adopted.

    Techniques: Plasmid Preparation, Clone Assay, Expressing, Western Blot, Concentration Assay

    Primer extension and northern blot analysis of thiC mRNA during meningococcal growth. (A) In order to identify the transcriptional start site of thiC (marked with red asterisks and arrow), primer extension reaction was performed with RNA isolated from N. meningitidis MC58 (wt) and ΔRS mutant, in which only the RS region upstream thiC is removed and therefore lacking the region where the radiolabelled primer anneals. ‘No RNA’ is the control reaction without RNA. (B) A schematic figure of the neisserial thiC gene. The location of both Northern blot probes was designed to anneal either to the thiC 5´UTR (probe A, red) or to the protein-coding region (probe B, blue). (C) RNA was isolated from N. meningitidis MC58 wt, a thiC (including the promoter and 5´UTR) knock out mutant (Δ thiC ) and a ΔRS mutant. Northern blot detection of thiC mRNA with probe A shows two RNA transcripts. A short fragment of about 190 nt is highly abundant at low OD 600 and the signal intensity decreases with the growth. The full-length mRNA (2200 nt) signal appears only at high OD 600 . The short fragment is not detected with probe B, indicating that it corresponds to the terminated 5´UTR. As expected, the mRNA is not present in the Δ thiC strain. In the case of ΔRS strain, probe B identifies the full-length mRNA, and its expression is independent of the growth phase.

    Journal: RNA Biology

    Article Title: Characterization of a transcriptional TPP riboswitch in the human pathogen Neisseria meningitidis

    doi: 10.1080/15476286.2020.1727188

    Figure Lengend Snippet: Primer extension and northern blot analysis of thiC mRNA during meningococcal growth. (A) In order to identify the transcriptional start site of thiC (marked with red asterisks and arrow), primer extension reaction was performed with RNA isolated from N. meningitidis MC58 (wt) and ΔRS mutant, in which only the RS region upstream thiC is removed and therefore lacking the region where the radiolabelled primer anneals. ‘No RNA’ is the control reaction without RNA. (B) A schematic figure of the neisserial thiC gene. The location of both Northern blot probes was designed to anneal either to the thiC 5´UTR (probe A, red) or to the protein-coding region (probe B, blue). (C) RNA was isolated from N. meningitidis MC58 wt, a thiC (including the promoter and 5´UTR) knock out mutant (Δ thiC ) and a ΔRS mutant. Northern blot detection of thiC mRNA with probe A shows two RNA transcripts. A short fragment of about 190 nt is highly abundant at low OD 600 and the signal intensity decreases with the growth. The full-length mRNA (2200 nt) signal appears only at high OD 600 . The short fragment is not detected with probe B, indicating that it corresponds to the terminated 5´UTR. As expected, the mRNA is not present in the Δ thiC strain. In the case of ΔRS strain, probe B identifies the full-length mRNA, and its expression is independent of the growth phase.

    Article Snippet: In order to assess the effect of different thiamine sources on the TPP-mediated expression of thiC in E. coli reporter strains and N. meningitidis , the following experimental procedures were adopted.

    Techniques: Northern Blot, Isolation, Mutagenesis, Knock-Out, Expressing

    Modulation of thiC expression in different growth media and bacterial or cell lysates. (A) E. coli carrying the reporter plasmid with EGFP fused to the wild-type N. meningitidis thiC promoter plus 5´UTR (pRS) or the variant with the mutation in the TPP binding region (pRS(BS)) was cultured in fresh or spent LB up to OD 600 = 0.1 and supplemented with H 2 O, spent LB, E. coli lysate and pure thiamine. After 2 and 4 h, bacterial samples were harvested and analysed by Western blot. EGFP signals were quantified and normalized to the corresponding RecA signals and each set by the value of the OD = 0.1 sample. (B) N. meningitidis was grown in either fresh or spent BHI supplemented with H 2 O, spent BHI, N. meningitidis lysate, A459 human alveolar epithelial cell lysate, and pure thiamine. Northern blot analysis was performed using probe A from Fig. 1 (B).

    Journal: RNA Biology

    Article Title: Characterization of a transcriptional TPP riboswitch in the human pathogen Neisseria meningitidis

    doi: 10.1080/15476286.2020.1727188

    Figure Lengend Snippet: Modulation of thiC expression in different growth media and bacterial or cell lysates. (A) E. coli carrying the reporter plasmid with EGFP fused to the wild-type N. meningitidis thiC promoter plus 5´UTR (pRS) or the variant with the mutation in the TPP binding region (pRS(BS)) was cultured in fresh or spent LB up to OD 600 = 0.1 and supplemented with H 2 O, spent LB, E. coli lysate and pure thiamine. After 2 and 4 h, bacterial samples were harvested and analysed by Western blot. EGFP signals were quantified and normalized to the corresponding RecA signals and each set by the value of the OD = 0.1 sample. (B) N. meningitidis was grown in either fresh or spent BHI supplemented with H 2 O, spent BHI, N. meningitidis lysate, A459 human alveolar epithelial cell lysate, and pure thiamine. Northern blot analysis was performed using probe A from Fig. 1 (B).

    Article Snippet: In order to assess the effect of different thiamine sources on the TPP-mediated expression of thiC in E. coli reporter strains and N. meningitidis , the following experimental procedures were adopted.

    Techniques: Expressing, Plasmid Preparation, Variant Assay, Mutagenesis, Binding Assay, Cell Culture, Western Blot, Northern Blot

    Structural analysis and ectopic reporter gene study of the cytX and thiD 5´UTR. (A) The predicted secondary structures of cytX TPP RS and thiD 5´UTRs (with RNAfold) are displayed. RBS and translational start codon (AUG) are depicted in red and orange, respectively. The alternative structure that could liberate the RBS of cytX while TPP is scarce is highlighted in blue. (B) The region corresponding to the promoter, 5´UTR and first 15 coding bp of cytX or 30 coding bp of thiD from N. meningitidis serogroups B were cloned in a translational fusion with EGFP. The expression of EGFP in E. coli was analysed by Western blot, in full (LB) and spent rich medium (spent LB), supplemented with thiamine. EGFP signals were quantified and normalized to the corresponding RecA signals and each set by the value of the LB 0 μM thiamine sample.

    Journal: RNA Biology

    Article Title: Characterization of a transcriptional TPP riboswitch in the human pathogen Neisseria meningitidis

    doi: 10.1080/15476286.2020.1727188

    Figure Lengend Snippet: Structural analysis and ectopic reporter gene study of the cytX and thiD 5´UTR. (A) The predicted secondary structures of cytX TPP RS and thiD 5´UTRs (with RNAfold) are displayed. RBS and translational start codon (AUG) are depicted in red and orange, respectively. The alternative structure that could liberate the RBS of cytX while TPP is scarce is highlighted in blue. (B) The region corresponding to the promoter, 5´UTR and first 15 coding bp of cytX or 30 coding bp of thiD from N. meningitidis serogroups B were cloned in a translational fusion with EGFP. The expression of EGFP in E. coli was analysed by Western blot, in full (LB) and spent rich medium (spent LB), supplemented with thiamine. EGFP signals were quantified and normalized to the corresponding RecA signals and each set by the value of the LB 0 μM thiamine sample.

    Article Snippet: In order to assess the effect of different thiamine sources on the TPP-mediated expression of thiC in E. coli reporter strains and N. meningitidis , the following experimental procedures were adopted.

    Techniques: Clone Assay, Expressing, Western Blot

    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. meningitidis with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.

    Journal: Biosensors & bioelectronics

    Article Title: Fully 3D Printed Integrated Reactor Array for Point-of-Care Molecular Diagnostics

    doi: 10.1016/j.bios.2018.03.009

    Figure Lengend Snippet: Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. meningitidis with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.

    Article Snippet: N. meningitidis was grown in Tryptic Soy Broth (BD, Sparks, MD) and counted on Trypticase™ Soy Agar with 5% Sheep Blood (BD, Sparks, MD).

    Techniques: Fluorescence, Lamp Assay, Chromatin Immunoprecipitation, Amplification, Concentration Assay

    Immunofluorescence microscopy of WCs of strains MC58, MC58Lnt and MC58LntC. To confirm the presence of meningococcal cells, cells were incubated with FITC‐labelled rabbit polyclonal IgG raised against WC N. meningitidis (left and right panel). To compare FHbp cell surface expression between the strains, cells were also incubated with anti‐FHbp antibody JAR4 that was detected by Alexa Fluor 555 labelled donkey anti‐mouse IgG secondary antibody (right panel).

    Journal: British Journal of Pharmacology

    Article Title: The role of apolipoprotein N‐acyl transferase, Lnt, in the lipidation of factor H binding protein of Neisseria meningitidis strain MC58 and its potential as a drug target) The role of apolipoprotein N‐acyl transferase, Lnt, in the lipidation of factor H binding protein of Neisseria meningitidis strain MC58 and its potential as a drug target

    doi: 10.1111/bph.13660

    Figure Lengend Snippet: Immunofluorescence microscopy of WCs of strains MC58, MC58Lnt and MC58LntC. To confirm the presence of meningococcal cells, cells were incubated with FITC‐labelled rabbit polyclonal IgG raised against WC N. meningitidis (left and right panel). To compare FHbp cell surface expression between the strains, cells were also incubated with anti‐FHbp antibody JAR4 that was detected by Alexa Fluor 555 labelled donkey anti‐mouse IgG secondary antibody (right panel).

    Article Snippet: The wells were washed three times in PBS then PBS with 1% ( w / v ) BSA containing 1:500 dilution Alexa Fluor 555 labelled donkey anti‐mouse IgG secondary antibody (Abcam) was added (to fluorescently label the FHbp‐bound JAR4 antibody) and 1:500 dilution of FITC‐labelled rabbit polyclonal IgG raised against WC N. meningitidis (Abcam) to detect meningococcal cells.

    Techniques: Immunofluorescence, Microscopy, Incubation, Expressing

    Dynamic meningococcal disease progression selects for Opa expression. (A) Mice with mild-disease pattern were imaged over 4 days using a detection scale of 200/2000 counts. Bioluminescent signals from the mice were undetectable at 2 days (2d) post-infection. From day 3 (3d), the signals reappeared either in the brain (upper panel) or in the whole body (lower panel). Images are representative of each subgroup. Each panel follows one mouse over time. Bacterial loads in blood and CSF at indicated time points post-challenge are shown. (B) N. meningitidis was isolated from blood, CSF or nasal washes (N wash) at day 1, 3 and 4 after infection. Recovered isolates together with the initial inoculum, were analyzed for expression of Opa proteins (30 kDa), PilC (110 kDa), and pili (PilE subunit 16 kDa) by immunoblots. LOS (6.5 kDa) was analyzed by Tricine SDS-PAGE.

    Journal: PLoS ONE

    Article Title: Imaging of Disease Dynamics during Meningococcal Sepsis

    doi: 10.1371/journal.pone.0000241

    Figure Lengend Snippet: Dynamic meningococcal disease progression selects for Opa expression. (A) Mice with mild-disease pattern were imaged over 4 days using a detection scale of 200/2000 counts. Bioluminescent signals from the mice were undetectable at 2 days (2d) post-infection. From day 3 (3d), the signals reappeared either in the brain (upper panel) or in the whole body (lower panel). Images are representative of each subgroup. Each panel follows one mouse over time. Bacterial loads in blood and CSF at indicated time points post-challenge are shown. (B) N. meningitidis was isolated from blood, CSF or nasal washes (N wash) at day 1, 3 and 4 after infection. Recovered isolates together with the initial inoculum, were analyzed for expression of Opa proteins (30 kDa), PilC (110 kDa), and pili (PilE subunit 16 kDa) by immunoblots. LOS (6.5 kDa) was analyzed by Tricine SDS-PAGE.

    Article Snippet: For detection of meningococci the tissue sections were incubated with rabbit antibody against N. meningitidis (diluted 1∶50; USBiological) for 60 min, horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (diluted 1∶50; BioRad) for 60 min, and stained using ImmunoPure metal Enhanced DAB substrate (Pierce).

    Techniques: Expressing, Mouse Assay, Infection, Isolation, Western Blot, SDS Page

    Translocation of N. meningitidis to the respiratory mucosa is dependent on PilC1. (A) Bioluminescent imaging of mice and nasal washes spread on GC-plates 24 h post infection. CD46 transgenic mice were infected i.v. with wild-type FAM20 LU or the PilC1-deficient mutant FAM20Δ pilC1 LU . Nontransgenic mice were challenged with FAM20 LU . Bacterial counts in the nasal washes (CFU/ml) are presented. Significantly fewer PilC1 mutants were recovered compared with wild-type bacteria ( P

    Journal: PLoS ONE

    Article Title: Imaging of Disease Dynamics during Meningococcal Sepsis

    doi: 10.1371/journal.pone.0000241

    Figure Lengend Snippet: Translocation of N. meningitidis to the respiratory mucosa is dependent on PilC1. (A) Bioluminescent imaging of mice and nasal washes spread on GC-plates 24 h post infection. CD46 transgenic mice were infected i.v. with wild-type FAM20 LU or the PilC1-deficient mutant FAM20Δ pilC1 LU . Nontransgenic mice were challenged with FAM20 LU . Bacterial counts in the nasal washes (CFU/ml) are presented. Significantly fewer PilC1 mutants were recovered compared with wild-type bacteria ( P

    Article Snippet: For detection of meningococci the tissue sections were incubated with rabbit antibody against N. meningitidis (diluted 1∶50; USBiological) for 60 min, horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (diluted 1∶50; BioRad) for 60 min, and stained using ImmunoPure metal Enhanced DAB substrate (Pierce).

    Techniques: Translocation Assay, Imaging, Mouse Assay, Infection, Transgenic Assay, Mutagenesis

    N. meningitidis targets the thyroid gland and the nasal area of CD46 transgenic mice. (A) BLI of mice at 24 h after i.v. infection. Distinct bioluminescent signals are visible in the thyroid area (T) and oral-nasal region (N) of CD46 transgenic mice. (B) Ex vivo BLI imaging of a dissected thyroid. (C) Bacterial loads in blood and thyroid at 24 h post-infection. Significant difference between thyroid of CD46 transgenic mice and nontransgenic mice is indicated ( P

    Journal: PLoS ONE

    Article Title: Imaging of Disease Dynamics during Meningococcal Sepsis

    doi: 10.1371/journal.pone.0000241

    Figure Lengend Snippet: N. meningitidis targets the thyroid gland and the nasal area of CD46 transgenic mice. (A) BLI of mice at 24 h after i.v. infection. Distinct bioluminescent signals are visible in the thyroid area (T) and oral-nasal region (N) of CD46 transgenic mice. (B) Ex vivo BLI imaging of a dissected thyroid. (C) Bacterial loads in blood and thyroid at 24 h post-infection. Significant difference between thyroid of CD46 transgenic mice and nontransgenic mice is indicated ( P

    Article Snippet: For detection of meningococci the tissue sections were incubated with rabbit antibody against N. meningitidis (diluted 1∶50; USBiological) for 60 min, horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (diluted 1∶50; BioRad) for 60 min, and stained using ImmunoPure metal Enhanced DAB substrate (Pierce).

    Techniques: Transgenic Assay, Mouse Assay, Infection, Ex Vivo, Imaging

    Generation and characterization of the bioluminescent N. meningitidis strain FAM20 LU . (A) Chromosomal integration of a bioluminescence expression cassette into strain FAM20. The luxCDABE operon of plasmid pLKMp is flanked by the Neisseria specific porA promoter (P) and terminator (T) sequences of the gapdh gene. The kanamycin resistance gene, kanR , enables selection on agar-plates. UHS and DHS represent two sequences homologous to a non-coding region between orf73 and orf74 of the FAM20 genome. (B) The porA promoter enhances bioluminescent emission. Bioluminescence (in photon units) emitted from N. meningitidis strain FAM20 transformed with pLKMp (black line) or transformed with control vector without promoter (grey line) is plotted against bacterial counts. (C) In vivo BLI of mice challenged i.p. with 10 8 FAM20 LU . Mice were imaged at indicated time points post-infection. In contrast to nontransgenic mice (upper panel), strong signals in the body together with focal signals in the brain were detected in CD46 transgenic mice (lower panel). Images show one representative mouse from each group.

    Journal: PLoS ONE

    Article Title: Imaging of Disease Dynamics during Meningococcal Sepsis

    doi: 10.1371/journal.pone.0000241

    Figure Lengend Snippet: Generation and characterization of the bioluminescent N. meningitidis strain FAM20 LU . (A) Chromosomal integration of a bioluminescence expression cassette into strain FAM20. The luxCDABE operon of plasmid pLKMp is flanked by the Neisseria specific porA promoter (P) and terminator (T) sequences of the gapdh gene. The kanamycin resistance gene, kanR , enables selection on agar-plates. UHS and DHS represent two sequences homologous to a non-coding region between orf73 and orf74 of the FAM20 genome. (B) The porA promoter enhances bioluminescent emission. Bioluminescence (in photon units) emitted from N. meningitidis strain FAM20 transformed with pLKMp (black line) or transformed with control vector without promoter (grey line) is plotted against bacterial counts. (C) In vivo BLI of mice challenged i.p. with 10 8 FAM20 LU . Mice were imaged at indicated time points post-infection. In contrast to nontransgenic mice (upper panel), strong signals in the body together with focal signals in the brain were detected in CD46 transgenic mice (lower panel). Images show one representative mouse from each group.

    Article Snippet: For detection of meningococci the tissue sections were incubated with rabbit antibody against N. meningitidis (diluted 1∶50; USBiological) for 60 min, horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (diluted 1∶50; BioRad) for 60 min, and stained using ImmunoPure metal Enhanced DAB substrate (Pierce).

    Techniques: Expressing, Plasmid Preparation, Selection, Transformation Assay, In Vivo, Mouse Assay, Infection, Transgenic Assay

    Bactericidal activity of protein A–purified mAb Me-1 against N. meningitidis strain 608B. Guinea pig serum ( closed circles ) or heat inactivated guinea pig serum ( open circles ) were used as the source of complement to evaluate the bactericidal activity.

    Journal: The Journal of Experimental Medicine

    Article Title: Highly Conserved Neisseria meningitidis Surface Protein Confers Protection against Experimental Infection

    doi:

    Figure Lengend Snippet: Bactericidal activity of protein A–purified mAb Me-1 against N. meningitidis strain 608B. Guinea pig serum ( closed circles ) or heat inactivated guinea pig serum ( open circles ) were used as the source of complement to evaluate the bactericidal activity.

    Article Snippet: A λGEM-11 genomic DNA library from N. meningitidis strain 608B was constructed according to the manufacturer's recommendations ( Promega , Madison, WI).

    Techniques: Activity Assay, Purification

    Nucleotide and amino acid sequences of the gene encoding the N. meningitidis strain 608B NspA protein. The open reading frame of 525 bp extends from the start codon at base 143 to the stop codon at base 667. A 19–amino acid leader peptide is underlined, and a putative ribosome binding site is boxed. These sequence data are available from GenBank under accession number U52066.

    Journal: The Journal of Experimental Medicine

    Article Title: Highly Conserved Neisseria meningitidis Surface Protein Confers Protection against Experimental Infection

    doi:

    Figure Lengend Snippet: Nucleotide and amino acid sequences of the gene encoding the N. meningitidis strain 608B NspA protein. The open reading frame of 525 bp extends from the start codon at base 143 to the stop codon at base 667. A 19–amino acid leader peptide is underlined, and a putative ribosome binding site is boxed. These sequence data are available from GenBank under accession number U52066.

    Article Snippet: A λGEM-11 genomic DNA library from N. meningitidis strain 608B was constructed according to the manufacturer's recommendations ( Promega , Madison, WI).

    Techniques: Binding Assay, Sequencing