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Lipopolysaccharides (LPS) ELISA Kit ELISA Kit

Reactivity: Various Species Colorimetric Competition ELISA 12.35 ng/mL - 1000 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Pubmed (22) Independent Validation (1)
Catalog No. ABIN6574100
$947.37
Plus shipping costs $45.00
96 tests
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  • Target
    Lipopolysaccharides (LPS)
    Reactivity
    • 3
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Various Species
    Detection Method
    Colorimetric
    Method Type
    Competition ELISA
    Detection Range
    12.35 ng/mL - 1000 ng/mL
    Minimum Detection Limit
    12.35 ng/mL
    Application
    ELISA
    Purpose
    The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of lipopolysaccharide in serum, plasma, tissue homogenates, cell lysates, cell culture supernates.

    We offer validation data (WB) for the kit components. So you can be sure to order a reliable ELISA kit product composed of high quality reagents.
    Sample Type
    Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
    Analytical Method
    Quantitative
    Specificity
    This assay has high sensitivity and excellent specificity for detection of Lipopolysaccharide (LPS)
    Cross-Reactivity (Details)
    No significant cross-reactivity or interference between Lipopolysaccharide (LPS) and analogues was observed.
    Sensitivity
    5.15 ng/mL
    Components
    • Pre-coated, ready to use 96-well strip plate, flat buttom
    • Plate sealer for 96 wells
    • Reference Standard
    • Standard Diluent
    • Detection Reagent A
    • Detection Reagent B
    • Assay Diluent A
    • Assay Diluent B
    • Reagent Diluent (if Detection Reagent is lyophilized)
    • TMB Substrate
    • Stop Solution
    • Wash Buffer (30 x concentrate)
    • Instruction manual
    Featured
    Discover our best selling ELISA Kit
  • Comment

    Information on standard material:
    The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

    Information on reagents:
    The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

    Information on antibodies:
    The provided antibodies and their host vary in different kits.

    Sample Volume
    50 μL
    Assay Time
    2 h
    Plate
    Pre-coated
    Protocol
    1. Prepare all reagents, samples and standards,
    2. Add 50μL standard or sample to each well.
      Then add 50μL prepared Detection Reagent A immediately.
      Shake and mix. Incubate 1 hour at 37 °C,
    3. Aspirate and wash 3 times,
    4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
    5. Aspirate and wash 5 times,
    6. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
    7. Add 50μL Stop Solution. Read at 450 nm immediately.
    Reagent Preparation
    1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
    2. Standard - Reconstitute the Standard with 0.5 mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 1,000 ng/mL. Please prepare 5 tubes containing 0.6 mL Standard Diluent and produce a triple dilution series according to the picture shown below. Mix each tube thoroughly before the next transfer. Set up 5 points of diluted standard such as 1,000 ng/mL, 333.33 ng/mL, 111.11 ng/mL, 37.04 ng/mL, 12.35 ng/mL, and the last EP tubes with Standard Diluent is the blank as 0 ng/mL.
    3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, kept for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
    4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
    5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

    Note:

    1. Making serial dilution in the wells directly is not permitted.
    2. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
    3. Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
    4. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
    5. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
    6. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
    7. Contaminated water or container for reagent preparation will influence the detection result.
    Sample Preparation
    • It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
    • If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
    • If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
    • Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).
    Assay Precision
    Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
    CV(%) = SD/meanX100
    Intra-Assay: CV < 10%
    Inter-Assay: CV < 12%
    Restrictions
    For Research Use only
  • Validation #103828 (ELISA)
    'Independent Validation' Badge
    by
    Nephrology Laboratory, Rush University Medical Center
    No.
    #103828
    Date
    05/01/2019
    Antigen
    LPS
    Lot Number
    L190202120
    Method validated
    ELISA
    Positive Control

    Serum from mice treated with LPS

    Negative Control

    Naïve serum samples from wt mice and mice knocked-out for gene relevant for our research

    Notes

    Passed, the LPS ELISA kit ABIN6574100 specifically detects LPS in mice serum.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    Secondary Antibody
    Full Protocol
    • Treat wt and ko mouse with LPS by intraperitoneal injection.
    • Collect the urine and serum 1h, 3h, and 6h after LPS injection.
    • Prepare all reagents and standards according to the manufacturer’s protocol.
    • Dilute samples 10x.
    • Add 50μl standard or sample to each well.
    • Add 50μl prepared Detection Reagent A immediately.
    • Shake and mix.
    • Cover plate with plate sealer and incubate for 1h at 37 °C.
    • Aspirate and wash 3x with 350µl 1x Wash Solution.
    • Add 100μl prepared Detection Reagent B.
    • Cover plate with plate sealer and Incubate for 30min at 37°C.
    • Aspirate and wash 5x with 350µl 1x Wash Solution.
    • Add 90μL Substrate Solution.
    • Cover plate with plate sealer and incubate 10min at 37°C.
    • Add 50μL Stop Solution and read plate at 450nm immediately.
    Experimental Notes
    • LPS in urine is undetectable even though the spike controls work well in urine. Possibly, the 1:10 dilution of the urine samples caused the LPS concentration to fall below the kit's detection limit. However, it was not possible to run undiluted urine samples because the mice got sick and had less urine after LPS treatment.

    • When measuring LPS in urine samples the LPS ELISA kit ABIN6574100 proved to be superior to the Endotoxin Assay Kit ABIN491527. Unlike ABIN6574100, ABIN491527 did not work with the spike control in urine.

  • Precaution of Use
    The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
    Storage
    4 °C/-20 °C
    Storage Comment
    1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
    2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.
    .
    Expiry Date
    6 months
  • Chen, Ji, Wang, Liao, Liang, Tang, Wen, Dominique, Li: "Involvement of TLR4 signaling regulated-COX2/PGE2 axis in liver fibrosis induced by Schistosoma japonicum infection." in: Parasites & vectors, Vol. 14, Issue 1, pp. 279, 2021 (PubMed).

    Kiang, Smith, Cannon, Anderson, Ho, Zhai, Cui, Xiao: "Ghrelin, a novel therapy, corrects cytokine and NF-κB-AKT-MAPK network and mitigates intestinal injury induced by combined radiation and skin-wound trauma." in: Cell & bioscience, Vol. 10, pp. 63, 2020 (PubMed).

    Sánchez-Tapia, Aguilar-López, Pérez-Cruz, Pichardo-Ontiveros, Wang, Donovan, Tovar, Torres: "Nopal (Opuntia ficus indica) protects from metabolic endotoxemia by modifying gut microbiota in obese rats fed high fat/sucrose diet." in: Scientific reports, Vol. 7, Issue 1, pp. 4716, 2019 (PubMed).

    Chen, Shen, Wang, Ding, Zhao, Wang, Fu, Wang: "Megasphaera elsdenii Lactate Degradation Pattern Shifts in Rumen Acidosis Models." in: Frontiers in microbiology, Vol. 10, pp. 162, 2019 (PubMed).

    Bluemel, Wang, Martino, Lee, Wang, Williams, Horvath, Stadlbauer, Zengler, Schnabl: "The Role of Intestinal C-type Regenerating Islet Derived-3 Lectins for Nonalcoholic Steatohepatitis." in: Hepatology communications, Vol. 2, Issue 4, pp. 393-406, 2018 (PubMed).

    Feng, Wang, Zhang, Gao, Tao, Ge, Zhu, Bi: "Modulation of gut microbiota contributes to curcumin-mediated attenuation of hepatic steatosis in rats." in: Biochimica et biophysica acta, Vol. 1861, Issue 7, pp. 1801-1812, 2017 (PubMed).

    Rosas-Villegas, Sánchez-Tapia, Avila-Nava, Ramírez, Tovar, Torres: "Differential Effect of Sucrose and Fructose in Combination with a High Fat Diet on Intestinal Microbiota and Kidney Oxidative Stress." in: Nutrients, Vol. 9, Issue 4, 2017 (PubMed).

    Hartmann, Seebauer, Mazagova, Horvath, Wang, Llorente, Varki, Brandl, Ho, Schnabl: "Deficiency of intestinal mucin-2 protects mice from diet-induced fatty liver disease and obesity." in: American journal of physiology. Gastrointestinal and liver physiology, Vol. 310, Issue 5, pp. G310-22, 2016 (PubMed).

    Wosiewicz, Żorniak, Hartleb, Barański, Hartleb, Onyszczuk, Pilch-Kowalczyk, Kyrcz-Krzemień: "Portal vein thrombosis in cirrhosis is not associated with intestinal barrier disruption or increased platelet aggregability." in: Clinics and research in hepatology and gastroenterology, 2016 (PubMed).

    Gao, Wen, Tong, Wang, Yang, Tang, Yan, Tai, Ye, Liu, Huang, Tang, Yang, Tang: "Inhibition of cyclooxygenase-2 alleviates liver cirrhosis via improvement of the dysfunctional gut-liver axis in rats." in: American journal of physiology. Gastrointestinal and liver physiology, Vol. 310, Issue 11, pp. G962-72, 2016 (PubMed).

    Zhao, Zhang, Chai, Li, Cui, Wang, Meng, Liu, Wang, Li, Bai, Xiao: "Oxymatrine attenuates CCl4-induced hepatic fibrosis via modulation of TLR4-dependent inflammatory and TGF-β1 signaling pathways." in: International immunopharmacology, Vol. 36, pp. 249-55, 2016 (PubMed).

    Avila-Nava, Noriega, Tovar, Granados, Perez-Cruz, Pedraza-Chaverri, Torres: "Food combination based on a pre-hispanic Mexican diet decreases metabolic and cognitive abnormalities and gut microbiota dysbiosis caused by a sucrose-enriched high-fat diet in rats." in: Molecular nutrition & food research, Vol. 61, Issue 1, 2016 (PubMed).

    Keshavarzi, Khaksari: "The effects of female sexual steroids on gastric function and barrier resistance of gastrointestinal tract following traumatic brain injury." in: Journal of pharmacy & bioallied sciences, Vol. 7, Issue 1, pp. 75-80, 2015 (PubMed).

    Karl, Fu, Wang, Zhao, Shen, Zhang, Wolfe, Saltzman, Zhao, Booth: "Fecal menaquinone profiles of overweight adults are associated with gut microbiota composition during a gut microbiota-targeted dietary intervention." in: The American journal of clinical nutrition, Vol. 102, Issue 1, pp. 84-93, 2015 (PubMed).

    Han, Suk, Kim, Kim, Baik, Kim, Cheon, Choi, Ham, Shin, Kim: "Effects of probiotics (cultured Lactobacillus subtilis/Streptococcus faecium) in the treatment of alcoholic hepatitis: randomized-controlled multicenter study." in: European journal of gastroenterology & hepatology, Vol. 27, Issue 11, pp. 1300-6, 2015 (PubMed).

    Barros, Borges, Ferreira, Carmo, Rosado, Fouque, Mafra: "Is there interaction between gut microbial profile and cardiovascular risk in chronic kidney disease patients?" in: Future microbiology, Vol. 10, Issue 4, pp. 517-26, 2015 (PubMed).

    Plaza-Diaz, Gomez-Llorente, Abadia-Molina, Saez-Lara, Campaña-Martin, Muñoz-Quezada, Romero, Gil, Fontana: "Effects of Lactobacillus paracasei CNCM I-4034, Bifidobacterium breve CNCM I-4035 and Lactobacillus rhamnosus CNCM I-4036 on hepatic steatosis in Zucker rats." in: PLoS ONE, Vol. 9, Issue 5, pp. e98401, 2014 (PubMed).

    Chen, Zhang, Dang, Liu, Tian, Zhao, Chen, Zhang, Chen: "Antidiabetic effect of Lactobacillus casei CCFM0412 on mice with type 2 diabetes induced by a high-fat diet and streptozotocin." in: Nutrition (Burbank, Los Angeles County, Calif.), Vol. 30, Issue 9, pp. 1061-8, 2014 (PubMed).

    He, Wang, Wu, Gong, Hong, Xia, Zhang, Bao, Gao: "Immunological activity difference between native calreticulin monomers and oligomers." in: PLoS ONE, Vol. 9, Issue 8, pp. e105502, 2014 (PubMed).

    Gram, Sveinsdottir, Ruscher, Hansson, Cinthio, Akerström, Ley: "Hemoglobin induces inflammation after preterm intraventricular hemorrhage by methemoglobin formation." in: Journal of neuroinflammation, Vol. 10, pp. 100, 2013 (PubMed).

  • Target
    Lipopolysaccharides (LPS)
    Alternative Name
    Lipopolysaccharide (LPS)
    Target Type
    Chemical
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