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LPS Extraction Kit

Cat.No Capacity Inquire
17141 100rxn Inquire
PRODUCT INFORMATION

* 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 Buffer100 ml x 1 ea
Purification Buffer

80 ml x 1 ea

 

Technical Data

 

 

Yield of LPS extraction

 

Strain 

LPS yield (μg)Protein contamination*
S. typhimurium200 ~ 400< 0.2 μg
S. enteritidis90 ~ 250< 0.2 μg
S. gallinarum150 ~ 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.

TroubleShooting Guide

Q. I am trying to pull out the LPS using the LPS kit. I want to use a 15ml tube because it has a lot of capacity, but the centrifuge that can use a 15ml tube has a maximum of 10,000rpm. The protocol is 10 minutes at 13,000rpm, but can I increase the time to 10,000 by 30 minutes?

 

A. In the case of our Extraction Kit, it is okay to perform centrifugation by extending the time by 20 to 30 minutes at 10,000rpm as mentioned. It is recommended in the protocol and centrifugation at 13,000 rpm for 10 minutes in order to check the optimal results. 

 

 

Q. I want to use Intron's LPS Extraction Kit, but can I try a sample?

 

A. Our LPS Extraction Kit was developed based on the phenol-water extraction method and is a product that can extract LPS easily and efficiently. We will take care of it as soon as possible by visiting the sales representative in charge.

 

Q. I want to isolate plant polysaccharides and see immune-related activities. I would like to know whether or not LPS (Lipopolysaccharide) is included in the plant polysaccharide. Are there any products that are kitted and can be easily identified?

 

A. plant does not have LPS, which we are talking about, which is present in bacteria. However, we know that there are various polysaccharides in plants. If you check the extraction of our LPS with silver stain, you can check it well.In other words, if you extract LPS with our LPS kit and silver stain, you can easily analyze the pattern of LPS.

 

Q. I am using the LPS extraction kit, but it seems that there is a little protein.

 

A. It seems to have been loaded. We guarantee the best results to use only the sample amount recommended by the protocol. In addition, if protein is present, it can be removed by treatment with Proteinase K. 

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