A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels

Tsai, Chao-Ming; Frasch, Carl E.

doi:10.1016/0003-2697(82)90673-x

Cited by 2,975 publications

(1,789 citation statements)

References 18 publications

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“…Lipooligosaccharide (LOS) bands were visualized by both silver staining and Alcian blue as described previously. 30,31 Static Light Scattering (SLS). SLS measurements were performed as reported.…”

Section: Methodsmentioning

confidence: 99%

A Unique Capsular Polysaccharide Structure from the Psychrophilic Marine Bacterium Colwellia psychrerythraea 34H That Mimics Antifreeze (Glyco)proteins

et al. 2015

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ABSTRACT:The low temperatures of polar regions and high altitude environments, especially icy habitats, present challenges for many microorganisms. Their ability to live under subfreezing conditions implies the production of compounds conferring cryotolerance. Colwellia psychrerythraea 34H, a -proteobacterium isolated from subzero Arctic marine sediments, provides a model for the study of life in cold environments. We report here the identification and detailed molecular primary and secondary structures of capsular polysaccharide from C. psychrerythraea 34H cells. The polymer was isolated in the water layer when cells were extracted by phenol/water and characterized by one-and two-dimensional NMR spectroscopy together with chemical analysis. Molecular mechanic and dynamic calculations were also performed. The polysaccharide consists of a tetrasaccharidic repeating unit containing two amino sugars and two uronic acids bearing threonine as substituent. The structural features of this unique polysaccharide resemble those present in antifreeze proteins and glycoproteins. These results suggest a possible correlation between the capsule structure and the ability of C. psychrerythraea to colonize subfreezing marine environments.

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Section: Methodsmentioning

confidence: 99%

A Unique Capsular Polysaccharide Structure from the Psychrophilic Marine Bacterium Colwellia psychrerythraea 34H That Mimics Antifreeze (Glyco)proteins

et al. 2015

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“…At the end of the purification, proteosomes (micellar multi-molecular organized structures) free of detergent were obtained [56] and will be referred to as "Nlac PorB". Purity of Nlac PorB was examined by Coomassie Brilliant blue staining of 12% SDS-PAGE [57] and silver stain of 15% SDS-PAGE [58]. Bacterial endotoxin content was determined with Limulus Amebocyte Lysate (LAL) assay as specified by the manufacturer (Cell Sciences, Canton, MA)…”

Section: Porin Purificationmentioning

confidence: 99%

The PorB porin from commensal Neisseria lactamica induces Th1 and Th2 immune responses to ovalbumin in mice and is a potential immune adjuvant

Liu

Wetzler

Massi

2008

Vaccine

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Porins from pathogenic Neisseriae are among several bacterial products with immune adjuvant activity. N. meningitidis (Nme) PorB, has been shown to induce immune cells activation in a TLR2-dependent manner and acts as a vaccine immune adjuvant. The PorB porin from Neisseria lactamica (Nlac), a common nasopharyngeal commensal shares significant structural and functional similarities with Nme PorB. In this work we ask whether the immune adjuvant ability of porins from pathogenic Neisserial strains is a characteristic shared with porins from non-pathogenic Neisserial species or whether it is unique for bacterial products derived from microorganisms capable of inducing inflammation and disease. We evaluate the potential immune adjuvant effect of Nlac PorB in mice using ovalbumin (OVA) as a prototype antigen. Immunization with Nlac PorB/OVA induced high OVA-specific IgG and IgM titers compared to OVA alone, similar to other adjuvants such as Nme PorB and alum. High titers of IgG1 and IgG2b were detected as well as production of IL-4, IL-10, IL-12 and INF-γ in response to Nlac PorB, consistent with induction of both a Th1-type and a Th2-type immune response. OVA-specific proliferation was also determined in splenocytes from Nlac PorB/OVA immunized mice. In addition, B cell activation in vitro and cytokine production in response to Nlac PorB was found to be mediated by TLR2, in a similar manner to Nme PorB.

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“…Electrophoresis was performed using a 350 Serological relationship between E. coli and Y. enterocolitica 351 constant current of 50 mAmp for 3-25 h. Profiles were either stained with silver [15,18] ELISA ELISAs were carried out as described previously [5,7]. Plates were coated with 06 ,ag of LPS and reacted with sera diluted (x 1000) in phosphate buffered saline (PBS) containing 05 % Tween-20 (PBS-Tween).…”

Section: Sds-pagementioning

confidence: 99%

The serological relationship betweenYersinia enterocoliticaO9 andEscherichia coliO157 using sera from patients with yersiniosis and haemolytic uraemic syndrome

Chart¹,

Cheasty²,

Cope

et al. 1991

Epidemiol. Infect.

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SUMMARYSera from patients with yersiniosis, shown to contain antibodies to Yersinia enterocolitica 09; and sera from patients with haemolytic uraemic syndrome (HUS) caused by Escherichia coli 0157, were used to investigate serological crossreactions between Y. enterocolitica 09 and E. coli 0157. Lipopolysaceharide (LPS) was isolated from strains of Y. enterocolitica 09 and E. coli 0157 and reacted with sera by immunoblotting and ELISA. Sera from patients with HUS contained antibodies to the LPS of E. coli 0157 only; 80 % of sera from patients with yersiniosis contained antibodies to the LPS of Y. enterocolitica 09 and E. coli 0157. This one-way cross-reaction was also detected using hyperimmune rabbit antisera.

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A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels

Cited by 2,975 publications

References 18 publications

A Unique Capsular Polysaccharide Structure from the Psychrophilic Marine Bacterium Colwellia psychrerythraea 34H That Mimics Antifreeze (Glyco)proteins

A Unique Capsular Polysaccharide Structure from the Psychrophilic Marine Bacterium Colwellia psychrerythraea 34H That Mimics Antifreeze (Glyco)proteins

The PorB porin from commensal Neisseria lactamica induces Th1 and Th2 immune responses to ovalbumin in mice and is a potential immune adjuvant

The serological relationship betweenYersinia enterocoliticaO9 andEscherichia coliO157 using sera from patients with yersiniosis and haemolytic uraemic syndrome

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