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IL1B Kit ELISA (Interleukin 1, beta) Kit ELISA

IL1B Reactivité: Humain AA 117-269 Colorimetric Sandwich ELISA 3.9-250 pg/mL Cell Culture Supernatant, Plasma (EDTA), Plasma (citrate), Plasma (heparin), Serum
Pubmed (31) Independent Validation (1)
N° du produit ABIN411293
$369.00
Plus shipping costs $45.00
96 tests
local_shipping Destination: Etats-Unis
Envoi sous 4 à 6 jours ouvrables
  • Antigène
    IL1B
    Épitope
    AA 117-269
    Reactivité
    Humain
    • 12
    • 9
    • 6
    • 5
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    3.9-250 pg/mL
    Seuil minimal de détection
    3.9 pg/mL
    Application
    ELISA
    Fonction
    Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human IL-1 beta
    Marque
    PicoKine™
    Type d'échantillon
    Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA), Plasma (citrate)
    Analytical Method
    Quantitative
    Specificité
    Expression system for standard: E.coli,A117-S269
    Réactivité croisée (Details)
    There is no detectable cross-reactivity with other relevant proteins.
    Sensibilité
    <0.15pg/mL
    Matériel non inclus
    Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl
    Immunogène
    Expression system for standard: E.coli
    Immunogen sequence: A117-S269
    Featured
    Discover our best selling IL1B Kit ELISA
  • Indications d'application
    Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.
    Commentaires

    Sequence similarities: Belongs to the IL-1 family.

    Plaque
    Pre-coated
    Protocole
    human IL-1 beta ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for IL-1 beta has been precoated onto 96-well plates. Standards (E.coli,A117-S269) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for IL-1 beta is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human IL-1 beta amount of sample captured in plate.
    Procédure de l'essai

    Aliquot 0.1 mL per well of the 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL, 16.5pg/mL, 7.8pg/mL, 3.9pg/mL human IL-1 betastandard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernatants, serum or plasma(heparin, EDTA, citrate) to each empty well. See "Sample Dilution Guideline" above for details. We recommend that each human IL-1 betastandard solution and each sample is measured in duplicate.

    Précision du teste
    • Sample 1: n=16, Mean(pg/ml): 19, Standard deviation: 1.0, CV(%): 5.3
    • Sample 2: n=16, Mean(pg/ml): 41.2, Standard deviation: 2.6, CV(%): 6.3
    • Sample 3: n=16, Mean(pg/ml): 116, Standard deviation: 6.5, CV(%): 5.6,
    • Sample 1: n=24, Mean(pg/ml): 27.5, Standard deviation: 1.6, CV(%): 5.8
    • Sample 2: n=24, Mean(pg/ml): 61.3, Standard deviation: 4.1, CV(%): 6.7
    • Sample 3: n=24, Mean(pg/ml): 179.1, Standard deviation: 12.7, CV(%): 7.1
    Restrictions
    For Research Use only
  • Validation #101868 (ELISA)
    'Independent Validation' signe
    by
    Kinderklinik, Universitätsklinikum, TU Dresden
    No.
    #101868
    Date
    16.05.2018
    Antigène
    IL1B
    Numéro du lot
    1141380914
    Application validée
    ELISA
    Contrôle positif
    Plasma from LPS stimulated human whole blood; Recombinant human IL-1B with increasing concentrations of canakinumab.
    Contrôle négative
    Performed following the instructions provided by the manufacturer.
    Conclusion

    Passed. ABIN411293 detects IL-1B in plasma from human whole blood. In the presence of increasing concentrations of canakinumab, false low IL-1B concentrations were detected, limiting the kit’s practicability.

    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    Anticorps secondaire
    Full Protocol
    • Collect blood from volunteers in hirudin coated tubes (Sarstedt, 04.1944.001).
    • Distribute blood samples on 96 well plates using 140µl per well.
    • Add ultra-pure LPS (Invivogen, tlrl-3pelps) to each well to a concentration of 1µg/ml to primer samples for NLRP3 assays.
    • Incubate plates on a shaker (450rpm) in a humidified incubator for 5.5h at 37°C and 5% CO2.
    • For NLRP3 inflammasome activation, add ATP (Invivogen, tlrl-atpl) to each well to a concentration of 1mM.
    • Incubate plates on a shaker (450rpm) in a humidified incubator for 30min at 37°C and 5% CO2.
    • Add 100µl PBS to each well.
    • Centrifuge plates for 5min at 1200rpm at RT and freeze the supernatant from each well at -80°C.
    • For analysis of canakinumab interference with ABIN411293, dilute recombinant IL-1B (BD, 558457) to 800pg/ml in RPMI1640 and incubate it with increasing doses of canakinumab (0µg/ml, 0.0001µg/ml, 0.001µg/ml, 0.01µg/ml, 0.1µg/ml, 1µg/ml, 10µg/ml, 100µg/ml) for 2h at RT.
    • Perform measurements according to the manufacturer’s protocol.
    • Analyze standards and samples in duplicate. Use assay buffer as blank and subtract the mean blank was subtracted from all raw data reads.
    • Use a five parameter logisitic curve to calculate the standard curve.
    Notes
    • The aim was to analyze the ability of ABIN411293 to detect IL-1B in plasma from stimulated human whole blood and in the presence of the therapeutic IL-1B antibody canakinumab.

    • Analysis of stimulated human whole blood showed the expected results, therefore ABIN411293 is suitable to detect IL-1B from plasma of stimulated human whole blood. Of note, increasing doses of canakinumab seem to interfere with the binding of the ELISAs antibodies leading to detection of false low IL-1B concentrations. Therefore, the ELISA may not be used to analyze serum or plasma from patients receiving therapeutic canakinumab.

  • Conseil sur la manipulation
    Avoid multiple freeze-thaw cycles.
    Stock
    -20 °C,4 °C
    Stockage commentaire
    Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles
    Date de péremption
    12 months
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    Wang, Qin, Wang, Chen, Lang, Zheng, Gao, Chen, Zhong, Mu, Wu, Zhang, Zhao, Zhong: "Pyroptosis induced by enterovirus 71 and coxsackievirus B3 infection affects viral replication and host response." dans: Scientific reports, Vol. 8, Issue 1, pp. 2887, 2018 (PubMed).

    Wan, Yuan, Liu, Xue: "miRNA-223-3p regulates NLRP3 to promote apoptosis and inhibit proliferation of hep3B cells." dans: Experimental and therapeutic medicine, Vol. 15, Issue 3, pp. 2429-2435, 2018 (PubMed).

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    Jian, Dai, Hu, Yao, Zheng, Zhu: "ox-LDL increases microRNA-29a transcription through upregulating YY1 and STAT1 in macrophages." dans: Cell biology international, Vol. 41, Issue 9, pp. 1001-1011, 2017 (PubMed).

    Huang, Liu, Ma, Liao, Lu, Huang, Qin, Liu, Fang: "Human cytomegalovirus triggers the assembly of AIM2 inflammasome in THP-1-derived macrophages." dans: Journal of medical virology, 2017 (PubMed).

    El-Horany, Abd-Ellatif, Watany, Hafez, Okda: "NLRP3 expression and urinary HSP72 in relation to biomarkers of inflammation and oxidative stress in diabetic nephropathy patients." dans: IUBMB life, Vol. 69, Issue 8, pp. 623-630, 2017 (PubMed).

    Sun, Zhu, Cai, Qiu: "Hypaphorine Attenuates Lipopolysaccharide-Induced Endothelial Inflammation via Regulation of TLR4 and PPAR-γ Dependent on PI3K/Akt/mTOR Signal Pathway." dans: International journal of molecular sciences, Vol. 18, Issue 4, 2017 (PubMed).

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    Wang, Lu, Zhang, Du, Tang, Wang, Lu, Hu: "GEN-27, a Newly Synthetic Isoflavonoid, Inhibits the Proliferation of Colon Cancer Cells in Inflammation Microenvironment by Suppressing NF-κB Pathway." dans: Mediators of inflammation, Vol. 2016, pp. 2853040, 2017 (PubMed).

    Alper, Erdogan, Erdogan, Bozan, Can: "Associations of Trauma Severity with Mean Platelet Volume and Levels of Systemic Inflammatory Markers (IL1β, IL6, TNFα, and CRP)." dans: Mediators of inflammation, Vol. 2016, pp. 9894716, 2017 (PubMed).

    Liu, Zhang, Ding, Zhou, Tao, Lu, Wang, Hu: "Nuclear Factor E2-Related Factor-2 Negatively Regulates NLRP3 Inflammasome Activity by Inhibiting Reactive Oxygen Species-Induced NLRP3 Priming." dans: Antioxidants & redox signaling, Vol. 26, Issue 1, pp. 28-43, 2017 (PubMed).

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  • Antigène
    IL1B
    Autre désignation
    IL1B (IL1B ELISA Kit Extrait)
    Synonymes
    IL-1, IL1-BETA, IL1F2, IL-1BETA, IL1beta, il1-b, zgc:111873, IL-1B, IL-1beta, Il-1b, IL1B, IL-1 beta, IL-1b, interleukin 1 beta, interleukin 1, beta, IL1B, il1b, Il1b
    Sujet

    Protein Function: Produced by activated macrophages, IL-1 stimulates thymocyte proliferation by inducing IL-2 release, B-cell maturation and proliferation, and fibroblast growth factor activity. IL-1 proteins are involved in the inflammatory response, being identified as endogenous pyrogens, and are reported to stimulate the release of prostaglandin and collagenase from synovial cells. .

    Background: Interleukin-1beta(IL-1beta) is a potent stimulator of bone resorption whose gene is mapped to 2q14, and has been implicated in the pathogenesis of high bone turnover and osteoporosis. IL-1beta, a prominent microglia-derived cytokine, caused oligodendrocyte death in coculture with astrocytes and microglia, but not in pure culture of oligodendrocytes alone. It also can cause nuclear export of a specific NCOR corepressor complex, resulting in derepression of a specific subset of nuclear factor-kappa-B(NFKB)-regulated genes. Furthermore, Microenvironmental IL-1beta and, to a lesser extent, IL-1alpha are required for in vivo angiogenesis and invasiveness of different tumor cells. Additional, the cooperation of IL-1beta and PDGFB induces contractile-to-synthetic phenotype modulation of human aortic smooth muscle cells in culture. Moreover, the association with disease may be explained by the biologic properties of IL-1beta, which is an important proinflammatory cytokine and a powerful inhibitor of gastric acid secretion.

    Synonyms: Interleukin-1 beta,IL-1 beta,Catabolin,IL1B,IL1F2,

    Full Gene Name: Interleukin-1 beta

    Cellular Localisation: Secreted. The lack of a specific hydrophobic segment in the precursor sequence suggests that IL-1 is released by damaged cells or is secreted by a mechanism differing from that used for other secretory proteins.
    ID gène
    3553
    UniProt
    P01584
    Pathways
    Signalisation NF-kappaB, Interferon-gamma Pathway, Signalisation TLR, Negative Regulation of Hormone Secretion, Cellular Response to Molecule of Bacterial Origin, Carbohydrate Homeostasis, Glycosaminoglycan Metabolic Process, Myometrial Relaxation and Contraction, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Autophagy, Cancer Immune Checkpoints, Inflammasome
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