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Lipopolysaccharides, from E. coli O55:B5

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MCE 國(guó)際站：Lipopolysaccharides, from E. coli O55:B5

產(chǎn)品活性：Lipopolysaccharides, from E. coli O55:B5 (LPS, from Escherichia coli (O55:B5)) 是從大腸桿菌 (E. coli O55:B5) 中提取的脂多糖內(nèi)毒素和 TLR-4 激活劑,，是一種 S (smooth) 型 LPS。Lipopolysaccharides, from E. coli O55:B5 具有典型的 3 部分結(jié)構(gòu)：O 抗原,、核心寡糖和脂質(zhì) A,。Lipopolysaccharides, from E. coli O55:B5 激活免疫細(xì)胞的 TLR-4，具有高致熱原性,，以及劑量和血清型特異性,。Lipopolysaccharides, from E. coli O55:B5 可用于誘導(dǎo)細(xì)胞炎癥和動(dòng)物炎癥相關(guān)模型。

研究領(lǐng)域：Immunology/Inflammation

作用靶點(diǎn)：Toll-like Receptor (TLR)

In Vitro: Note:
To maintain the integrity of LPS, it is recommended to store LPS solution in silanized containers. This is because LPS can adhere to plastics and certain types of glass, particularly at concentrations below 0.1 mg/mL. If the LPS concentration exceeds 1 mg/mL, this adsorption effect is relatively minimal. If using glass containers, ensure that the solution is thoroughly mixed for at least 30 minutes before use to redissolve any LPS that may have adsorbed to the tube walls.

LPS is the major toxic component of Gram-negative bacteria, capable of activating pathogen-associated molecular patterns (PAMP) of the immune system and inducing cellular secretion of migrasomes. LPS can be recognized by TLR4, activating the innate immune system, followed by the promotion of NF-κB activation and the production of pro-inflammatory cytokines. It is commonly used in experiments involving the stimulation, activation, and differentiation of immune cells.
Different bacterial species express LPS with varying structures and biological activities. LPS generally exists in two configurations: R (rough) type and S (smooth) type. S-type LPS contains a typical three-part structure: O-antigen (O-antigen) (serum-specific polysaccharides composed of repeating oligosaccharide units), core oligosaccharide (core) (non-repeating C9-type oligosaccharides), and lipid A (Lipid A) (the toxic component of LPS). The R type lacks the O-antigen and expresses rough-type LPS. The absence of O-antigen can affect the process of immune cell recognition of LPS.
The LPS expressed by the E. coli 055:B5 strain serves as a prototype endotoxin frequently used as an endotoxin standard in LAL assays. Lipopolysaccharides, from E. coli O55:B5 exhibit high pyrogenicity and are commonly used for in vitro cell activation. Lipopolysaccharides, from E. coli O55:B5 induce the secretion of pro-inflammatory cytokines in mouse macrophages.

1. Induction of cellular inflammation model

Background

LPS binds to the TLR4-MD-2 complex on the cell surface, activates both MyD88-dependent and MyD88-independent signaling pathways, promotes the translocation of transcription factors such as NF-κB into the nucleus, triggers the expression of inflammation-related genes, and leads to the occurrence of an inflammatory response in cells.

Specific Modeling Methods

Cell: Macrophages, tumor cells, glial cells and so on.
Administration: 0.1-10 μg/mL ? 1-24 h

Note

(1) Before the formal experiment, relevant references should be consulted according to the cell line and the source of LPS, etc., and concentration and time gradients should be screened to determine the optimal experimental protocol.
(2) Stimulating cells with LPS does not necessarily lead to cell death. Therefore, it is not appropriate to determine the concentration and time of LPS for establishing the model merely by detecting cell viability. It is recommended to detect the expression and secretion of multiple inflammatory factors.
(3) During the process of stimulating cells with LPS, the morphological changes of cells should be observed regularly. An excessively high concentration may cause cytotoxicity, while an excessively low concentration may fail to effectively damage the cells.
(4) A certain concentration of DMSO can significantly inhibit the inflammatory response induced by LPS. It is recommended to dissolve LPS in PBS or ddH₂O.
(5) In the research on the construction of in vitro inflammatory models, Lipopolysaccharides, from E. coli O55:B5 (HY-D1056) and Lipopolysaccharides, from E. coli O111:B4 (HY-D1056A1) are the most widely used LPS, and they are highly recommended!

Modeling Indicators

The secretion/expression of inflammatory factors such as IL-1β, IL-6 and TNF-α in the supernatant or cells increases.
The release of NO increases.
The expression of inflammatory genes such as iNOS, NF-κB and NLRP3 increases.

Correlated Product(s) Lipopolysaccharides, from E. coli O111:B4 (HY-D1056A1)

Opposite Product(s): /

In Vivo: Lipopolysaccharides, from E. coli O55:B5 can be used to induce cell and animal models related to inflammation. Among them, the animal models include sepsis (shock) model, cardiac dysfunction/myocarditis model, acute lung injury model, acute liver injury model, encephalitis model, depression model, etc. The following are examples of several models:

1. Induction of acute liver failure model

Background

LPS binds to the receptors on the cell surface, activates inflammatory cells to release a large amount of inflammatory mediators, and triggers an inflammatory response. The excessive activation of inflammation leads to lung tissue damage, pulmonary edema, and pulmonary dysfunction, thus establishing an acute lung injury model.

Specific Modeling Methods

C57/BALB/c mice: 0.2-15 mg/kg ? Intratracheal administration

Note

(1) Before inducing an animal model with LPS, relevant references should be consulted based on the experimental purpose, animal type, etc., and preliminary experiments should be conducted to determine the optimal experimental protocol.
(2) After LPS administration, the time points at which the peak levels of different inflammatory factors appear may vary. It is recommended to determine the experimental protocol according to references, and multiple time points should be selected for detection during preliminary experiments.
(3) LPS should be stored away from light and avoid repeated freezing and thawing.

Modeling Indicators

The secretion/expression of TNF-α, IL-6, IL-1β, etc. in the BALF or lung tissue decreases.
HE staining of lung tissue: Leukocyte infiltration in the alveoli, thickening of the alveolar wall, patchy hemorrhage, interstitial edema, etc.
The dry/wet ratio (D/W ratio) of the lung decreases.

Correlated Product(s): Lipopolysaccharides, from E. coli O111:B4 (HY-D1056A1)

Correlated Product(s): /

2. Induction of cardiac dysfunction/myocarditis model

Background

Cardiac dysfunction (myocarditis) is a common complication of sepsis induced by LPS. LPS can activate the inflammatory response and oxidative stress, causing damage and apoptosis of cardiomyocytes as well as cardiac fibrosis, which in turn leads to cardiac dysfunction.

Specific Modeling Methods

C57/ HsdWin:NMRI mice ? 10 mg/kg i.p.

Modeling Indicators

Myocardial dysfunction: The left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) decreases, and the left ventricular end-systolic volume (LVESV) and left ventricular end-systolic diameter (LVESD) increases.
The levels of CK-MB, LDH, AST, TNF-α, IL-6, IL-1β, etc. in the serum increases, and the TLR4/NF-κB pathway inhibites.
HE staining of the heart: The arrangement of myocardial fibers is disordered, the myocardial tissue is damaged with unclear contours, there are phenomena such as myocardial lysis, interstitial edema, congestion, infiltration of inflammatory cells, etc.

Correlated Product(s): Lipopolysaccharides, from E. coli O111:B4 (HY-D1056A1)

Opposite Product(s): /

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