The outer membrane of gram-negative bacteria contains lipopolysaccharide (LPS), a low molecular weight carbohydrate with a molecular mass of 10-20 kDa. It is heterogeneous, and composed of O antigen (a repeating glycan polymer), core oligosaccharide (which links the O antigen to Lipid A - the third component, and non-carbohydrate components such as phosphate and amino acids groups. Lipid A, has multiple fatty acids which serve to anchor LPS into the bacterial membrane allowing the O antigen and core oligosaccharide to protrude and contributes to a major part of the toxicity of gram-negative bacteria. Also known as endotoxin, when consumed by animals, LPS induces a strong inflammatory response and/or sepsis.
Firdous Hassan I think the Hot-Phenol & Proteinase K method are good.
Razieh Sadat Hosseini Thanks for immediate and kind response, actually from literature most people use mannual method but there are kit based protocol as well, what could be better option?. On earlier i have extracted LPS via mannual method through hot-phenol and proteinase K method @ DOI: 10.3791/3916 , but there are complications when running SDS PAGE, on final step blue layer is homogeneous when added 2XSDS buffer, but when i added 2xSDS buffer to aqueous clear layer within no it time SDS buffer settled down visualizing again two layers then how could i load the this sample.
Response would be highly appreciated..
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