Reagentia en instrumenten voor immunologie, celbiologie en moleculaire biologie.
 
> Lipopolysaccharides, from E. coli O55:B5

Lipopolysaccharides, from E. coli O55:B5

Afdrukken Afdrukken
Log in alstublieft

Referentie HY-D1056-5mg

Formaat : 5mg

Merk : MedChemExpress


Description

Lipopolysaccharides, from E. coli O55:B5 (LPS, from Escherichia coli (O55:B5)) are endotoxins and TLR4 activators extracted from Escherichia coli (E. coli O55:B5) and are classified as S (smooth) type LPS. Lipopolysaccharides, from E. coli O55:B5 possess the typical three-part structure: O-antigen, core oligosaccharide, and lipid A. Lipopolysaccharides, from E. coli O55:B5 activate TLR-4 in immune cells, exhibit high pyrogenicity, and demonstrate dose and serotype specificity. Lipopolysaccharides, from E. coli O55:B5 can be widely used to induce cellular inflammation and establish animal models related to inflammation[1][2][3][4][5][6][7].
It is recommended to prepare a solution with concentration ≥2 mg/mL. Vortex thoroughly for more than 10 minutes. Due to the adsorption characteristics of LPS, silanized container or low adsorption centrifuge tubes should be used for aliquoting and storage, and mix thoroughly before use.

IC50 & Target

TLR4

 

In Vitro

Note:
1. Concentration and Time: Please do not rely solely on a single article to determine experimental conditions. It is recommended to review relevant literature based on the cell line and type of LPS before formal experiments, as the required induction time or optimal concentration for different inflammatory factors to reach their peak may vary. It is advisable to set concentration and time gradients to identify the optimal experimental scheme.
2. Detection Indicators: LPS does not necessarily induce cell death; therefore, it is not appropriate to determine the LPS modeling concentration and time solely by assessing cell viability. It is recommended to measure the expression or secretion of inflammatory factors.
3. Solvent Selection: Literature indicates that certain concentrations of DMSO can significantly inhibit LPS-induced inflammatory responses. In cellular experiments, it is recommended to prepare stock solutions using sterile water, followed by dilution with culture medium.
4. Container Selection: Due to the adsorption characteristics of LPS, it can bind to plastics and certain types of glass (especially at concentrations <0.1 mg/mL). The adsorption effect is relatively small when LPS concentrations exceed 1 mg/mL. Additionally, LPS tends to form micelles in solution. Therefore, when dissolving the powder, it is recommended to prepare concentrations of ≥2 mg/mL, and to vortex thoroughly for more than 10 minutes. If necessary, ultrasonic assistance may be used. For storage, please use silanized containers or low-adhesion centrifuge tubes. If glass containers are used, ensure to mix thoroughly for at least 30 minutes prior to use to re-dissolve any LPS adsorbed to the wall of the container.
5. Concentration Units: LPS does not have a uniform molecular weight because its molecules exhibit heterogeneity and aggregation. The molecular weight of naturally sourced LPS typically ranges from 10-100 kDa or even higher. Common dosing concentrations for LPS found in the literature are in terms of mass concentration, such as ng/mL and μg/mL, so it is sufficient to prepare solutions directly in mass concentration during experiments.
6. Filtration Sterilization: After dissolving LPS powder in water, saline, or PBS, the solution may appear turbid or colloidal, and in some cases, a microsphere distribution with diameters around 20-30 nm may be observed. When sterilizing by filtration, do not filter the stock solution directly. It is recommended to dilute to working solution first and then filter sterilize through a 0.22 μm filter membrane.
7. Differences Among Different Strain LPS: LPS of different catalog numbers comes from various bacterial strains, corresponding to different structural features such as lipid A, core polysaccharides, and O-antigens, which in turn affect the intensity of inflammation induction and TLR4-mediated signaling bias. Commonly referenced LPS catalog numbers for in vitro or in vivo inflammation model construction include HY-D1056 and HY-D1056A1. Moreover, in specific research contexts, specialized sources of LPS related to the studied bacterial strains may also be used. For example, HY-D1056D (from Porphyromonas gingivalis) is used in periodontal studies, while HY-D1056B3 (from Klebsiella pneumoniae) is relevant in pneumonia-related research. When selecting LPS, considerations should include the purpose of the experiment, sensitivity of the cell line, and other factors.

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].

1. Induction of cellular inflammation model[4][5]
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)

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Lipopolysaccharides, from E. coli O55:B5 Related Antibodies
In Vivo

Note:
Please do not refer to only one article to determine the experimental conditions. It is recommended to determine the optimal experimental conditions (animal strain, age, dosage, frequency and cycle, detection time and indicators, etc.) through preliminary experiments before the formal experiment.

Lipopolysaccharides (1.5 mg/kg; i.p.; once) induces sickness and hypothermia in mice, and induces a greater and more prolonged sickness response in adult male mice[3].

Guidelines for Establishing In Vivo Animal Models with LPS

Animal Models Recommended LPS Types Reference Doses and Management Methods (Taking Mice as an Example) Recommended Detection Indicators
Sepsis (Shock) Model [PMID: 26440998; 27127234; 19529910] LPS, from E. coli O111:B4 (HY-D1056A1)
LPS, from E. coli O55:B5 (HY-D1056)
LPS, from E. coli O127:B8 (HY-D1056A2)		 2.5-10 mg/kg;
Intraperitoneal injection		 Assessment of Inflammatory Mediators: TNF-α, IL-6, IL-1β, etc. (ELISA detection of Serum/Plasma).
Histopathological Evaluation: HE staining of the kidney, glomerular cell proliferation, epithelial cell degeneration and necrosis, interstitial inflammatory cell infiltration, etc.
Others: Serum Scr, BUN (markers of renal filtration function).
Cardiac Dysfunction/Myocarditis Model [PMID: 32896106; 36593471] LPS, from E. coli O111:B4 (HY-D1056A1)
LPS, from E. coli O55:B5 (HY-D1056)		 10 mg/kg;
Intraperitoneal injection 		Assessment of Inflammatory Mediators: TNF-α, IL-6, IL-1β, etc. (ELISA detection of Serum/Plasma).
Histopathological Evaluation: HE staining of the heart, disordered arrangement of myocardial fibers, destruction of myocardial tissue, unclear contour, myocardial dissolution, interstitial edema, congestion, inflammatory cell infiltration, etc.
Others: Echocardiographic detection of dysfunction.
Acute Lung Injury Model [PMID: 31595149; 26888116; 20975550] LPS, from E. coli O111:B4 (HY-D1056A1)
LPS, from E. coli O55:B5 (HY-D1056)
LPS, from Klebsiella pneumonia (HY-D1056B3) 		0.2-15 mg/kg;
Intratracheal administration 		Assessment of Inflammatory Mediators: TNF-α, IL-6, IL-1β, etc. (ELISA/qPCR detection of BALF/lung tissue).
Histopathological Evaluation: HE staining of the Lung, leukocyte infiltration in alveoli, thickening of alveolar walls, plaque hemorrhage and interstitial edema, etc.
Others: Lung dry/wet ratio (D/W ratio).
Acute Liver Injury Model [PMID: 27127234; 36849063] LPS, from E. coli O111:B4 (HY-D1056A1)
LPS, from E. coli O55:B5 (HY-D1056) 		5-50 μg/kg + D-GalN (200-400 mg/kg);
Intraperitoneal injection		 Assessment of Inflammatory Mediators: TNF-α, IL-6, IL-1β, etc. (ELISA/qPCR detection of serum/liver tissue).
Histopathological Evaluation: HE staining of the liver, liver structural disorder, hemorrhagic plaques, inflammatory cell infiltration, etc.
Others: Serum ALT, AST (markers of liver injury).
Periodontitis Model [PMID: 23167849; 27987467] LPS, from P. gingivalis (HY-D1056D)
LPS, from E. coli O55:B5 (HY-D1056) 		4-10 μg; 2-4 times;
Oral periodontal tissue injection 		Assessment of Inflammatory Mediators: iNOS, COX-2, TNF-α, IL-6, etc. (qPCR detection of gingival tissue).
Histopathological Evaluation: HE staining of gingival/alveolar bone tissue, Inflammatory cells infiltrate, and collagen fiber bundles are loosely distributed near the tissue-root interface.
Others: The number of immune cells in gingival tissue (flow cytometry).
Encephalitis Model [PMID: 35858866; PMID: 36240654] LPS, from E. coli O111:B4 (HY-D1056A1)
LPS, from P. gingivalis (HY-D1056D) 		3-5 mg/kg;
Intraperitoneal injection		 Assessment of Inflammatory Mediators: IL-1β, IL-6, TNF-α, etc. (qPCR/WB detection of brain/hippocampus tissue).
Immunofluorescence Detection: IBA-1 (Microglia marker); GFAP (Astrocyte marker); Siglec-E, etc.
Depression Model [PMID: 31327964; 38677623] LPS, from E. coli O111:B4 (HY-D1056A1)
LPS, from E. coli O55:B5 (HY-D1056) 		5-10 mg/kg;
Intraperitoneal injection		 Assessment of Inflammatory Mediators: IL-1β, NLRP3, Caspase-1, etc. (qPCR/WB detection of hippocampus/cortex); IL-6, IL-1β, TNF-α, etc. (ELISA detection of hippocampus/cortex/serum).
Immunofluorescence Detection: IBA-1, GFAP, etc.
Behavioral Tests/Detection: Tail suspension experiments and forced swimming experiments, etc., has a longer time of immobility and a shortened latency period of immobility.
Parkinson's Model [PMID: 35065246; 30455692] LPS, from E. coli O111:B4 (HY-D1056A1)
LPS, from P. gingivalis (HY-D1056D) 		2-5 μg;
Black matter injection 		Assessment of Inflammatory Mediators: iNOS, COX-2, etc. ( WB detection of substantia nigra); TNF-α, IL-1β, IL-6, etc. (ELISA/qPCR detection of substantia nigra).
Immunofluorescence Assay/Detection: IBA-1, GFAP, etc.
Behavioral Tests/Detection: Rotational behavior test, significantly increased number of rotations; Open field test, bradykinesia, reduced exploratory drive in novel environment, preference for peripheral zone activity; Rotarod test, shortened retention time, premature fall latency.
Others: Cx43, Tyrosine hydroxylase (WB detection of substantia nigra).
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[6]
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[7]
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)

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model: Female and male CD1 mice[3]
Dosage: 1.5mg/kg
Administration: Intraperitoneal injection, once
Result: Induced sickness behavior in all mice, but adult mice displayed more sickness than pubertal mice and adult males remained sick for a longer period of time than adult females.
Caused a decrease in body temperature for all mice, but this decrease was greatest in adult males.
Increased pro- and anti-inflammatory cytokines at various levels in pubertal and adult male and female mice, resulted in age and sex differences in cytokine concentrations following immune challenge.
Only adult males and females treated with LPS displayed significantly more IL-6 than their saline controls, and pubertal males and females and adult females displayed significantly more IL-10 than their saline controls.
All the mice displayed significantly more IL-12 and TNF-α than their saline controls.
Essai clinique
Appearance

Solid

Color

White to off-white

SMILES

[Lipopolysaccharides, from E. coli O55:B5]
Structure Classification
Initial Source

surface of Gram-negative bacteria
Livraison

Room temperature in continental US; may vary elsewhere.
Stockage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 6 months
-20°C 1 month
Solvant et solubilité
In Vitro: 

H2O : 5 mg/mL (Need ultrasonic; DMSO can inactivate Lipopolysaccharides, from E. coli O55:B5's activity)

For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  PBS

    Solubility: 8.33 mg/mL; Clear solution; Need ultrasonic and warming and heat to 60°C

In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Calculation results:
Working solution concentration: mg/mL
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only.If necessary, please contact MedChemExpress (MCE).
Pureté et documentation
Références

Misschien heeft u ook interesse in de volgende producten:



Referentie
Beschrijving
Cond.
Price Bef. VAT
AAH-APO-1-2
Human Apoptosis Array C1
 2SampleKit 
AAH-APO-1-8
Human Apoptosis Array C1
 8SampleKit 
AAH-APO-1-4
Human Apoptosis Array C1
 4SampleKit