LPS Extraction Kit
* This product contains hazardous chemicals (phenols and mixtures containing more than 5% of them). When handling, please observe the handling standards (can be checked in MSDS).
Description
A product that can separate/purify LPS existing on the outer wall of Gram-negative bacteria
• Isolation/purification of LPS in Gram-negative bacteria
• Phenol-water method of LPS extraction
• Effective removal of contaminants such as proteins and nucleic acids
• The most efficient and simple LPS extraction product
• The entire process of extracting LPS is completed in up to 60 minutes
LPS Extraction Kit is a product that can separate/purify LPS existing on the outer cell wall of Gram-negative bacteria. In general, Gram-negative bacteria have a separate structure called lipopolysaccharide (LPS) in addition to phospholipid and protein in the external layer of the cell envelope, and this has different characteristics for each bacteria. Such LPS is known to play an important role in bacterial growth and survival, and especially in the interaction between host and parasite. In addition, LPS is understood to be closely related to the immune response, such as causing peripheral vascular collapse by secreting endotoxin when septic shock is applied, and is also important for toxic pathophysiological responses. The extraction pattern of LPS is different for each bacteria, and through this, the bacteria are systematically separated. As mentioned earlier, bacterial endotoxins are generally present on the outer walls of Gram-negative bacteria and are not expressed while the bacteria are alive, but are released to the outside by breaking the walls of the cells when the bacteria proliferate or die. Natural entotoxin is composed of LPS, Proteins, and Phospholipids. It is very stable in nature, resistant to heat, negatively charged, and has a large molecular weight (more than 1,000,000 daltons). In general, proteins are extracted with Trichloroacetic acid, Butanol or EDTA to remove lipid associated proteins (LAP), and phospholipids are extracted using phenol. Typically, if you try to extract LPS, you can get around 1-4% LPS per bacterial dry weight.Since the LPS Extraction Kit is based on the Phenol water method, it denatures the protein in entotoxin and removes it by denaturing it in the form of a precipitate between the aqueous layer and the phenol layer.Since the lipid is separated by the phenol/chloroform fraction, only high LPS can be extracted quickly and easily.
Applications
01 LPS composition and structure study
02 Phylogenetic study of bacteria
03 Antibiotic target research
04 LPS inhibitory drug design study
05 Carbohydrate antigen immune response study
Kit Contents
| Lysis Buffer | 100 ml x 1 ea |
| Purification Buffer | 80 ml x 1 ea |
Technical Data
Yield of LPS extraction
Strain | LPS yield (μg) | Protein contamination* |
|---|---|---|
| S. typhimurium | 200 ~ 400 | < 0.2 μg |
| S. enteritidis | 90 ~ 250 | < 0.2 μg |
| S. gallinarum | 150 ~ 450 | < 0.2 μg |
| E. coli (wild type) | 220 ~ 490 | < 0.2 μg |
| E. coli (O:055) | 260 ~ 510 | < 0.2 μg |
| E. coli (O:111) | 220 ~ 500 | < 0.2 μg |
| E. coli (O:1) | 180 ~ 380 | < 0.2 μg |
| E. coli (O:2) | 180 ~ 380 | < 0.2 μg |
5 After extracting LPS from each G(-) strain of OD600 using the LPS Extraction Kit, the extracted LPS was quantified using Purpald Assay, and as a result, the above extraction efficiency was confirmed. On the other hand, the presence of protein was observed using our SMART™ Micro BCA Assay Kit, and contamination of less than 0.2 μg was confirmed.
Band pattern of LPS extracted from various strains
Lane 1 : S. typhimurium Lane 2 : S. enteritidis
Lane 3 : E. coli (O055) Lane 4 : E. coli (O111)
Lane 5 : S. gallinarium Lane 6 : S. enterititis
Lane 7 : S. typhimurium Lane 8 : E. coli (wild tyoe)
Lane 9 : E, coli (O111) Lane 10 : E. coli (O2)
As a result of silver staining after SDS-PAGE of LPS extracted from various types of Gram-negative bacteria, a band pattern in the form of a ladder is observed according to the characteristics of the LPS present as a multimer and the number of multimers. It is observed that there is a difference in pattern.
Citation List
| 1 | Vaccine Volume 36, Issue 29, 5 July 2018, Pages 4153-4156 |
Immunization with lipopolysaccharide-free outer membrane complexes protects against Acinetobacter baumannii infection | Marina R. Pulido a, Meritxell García-Quintanilla a, Jerónimo Pachón a, b, Michael J. McConnell | Spain |
| 2 | bioRxiv, 2023 | Understanding the Mechanisms of Salmonella Typhimurium resistance to Cannabidiol | Iddrisu Ibrahim, View ORCID ProfileJoseph Atia Ayariga, Junhuan Xu, Daniel A. Abugri, Robertson K. Boakai, Olufemi S. Ajayi | USA |
| 3 | Biosensors 2022, 12, 153. https://doi.org/10.3390/bios12030153 https://www.mdpi.com/journal/biosensors | A Novel Peptide as a Specific and Selective Probe for Klebsiella pneumoniae Detection | Hyun Kim, Ju Hye Jang , Young Jung, Ju Hyun Cho | Korea |
| 4 | Int. J. Mol. Sci. 2022, 23, 5442. https://doi.org/10.3390/ijms23105442 https://www.mdpi.com/journal/ijms | Prunetinoside Inhibits Lipopolysaccharide-Provoked Inflammatory Response via Suppressing NF-κB and Activating the JNK-Mediated Signaling Pathway in RAW264.7 Macrophage Cells | Abuyaseer Abusaliya, Pritam Bhagwan Bhosale, Hun Hwan Kim, Sang Eun Ha, Min Yeong Park, Se Hyo Jeong , Preethi Vetrivel, Joon-Suk Park, Gon Sup Kim | Korea |
| 5 | Journal
of Oral Biosciences Available online 2 June 2022 |
Green tea catechins inhibit Porphyromonas gulae LPS-induced inflammatory responses in human gingival epithelial cells | ShoYoshida, HiroakiInaba, RyotaNomura, Kazuhiko Nakano, Michiyo Matsumoto-Nakano | Japan |
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