IL-1 beta ELISA Kit

Produktnummer ABIN411293

Dieses Colorimetric ELISA kit dient der quantitativen Messung von Human IL-1 beta und wurde unabhängig validiert. Es wurde in 31 Publikationen zitiert.

Kurzübersicht für IL-1 beta ELISA Kit (ABIN411293)

Target: IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))

Bindungsspezifität: AA 117-269

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 3.9 pg/mL - 250 pg/mL

Applikation: ELISA

Proben: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA), Plasma (citrate)

Produktdetails IL-1 beta ELISA Kit

Untere Nachweisgrenze: 3.9 pg/mL

Verwendungszweck: Sandwich Human IL-1 Beta/IL-1F2/IL1B ELISA Kit to quantitate Human IL1B in cell culture supernatants, serum and plasma (heparin, EDTA, citrate).

Analytische Methode: Quantitative

Spezifität: Expression system for standard: E.coli
Immunogen sequence: A117-S269

Capture antibody and Detection antibody: a monoclonal antibody from mouse, a biotinylated monoclonal antibody from mouse

Kreuzreaktivität (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivität: < 0.15 pg/mL

Bestandteile:

1. Pre-coated 96-well strip microplate
2. Standard
3. Biotinylated antibody (100x)
4. Avidin-Biotin-Peroxidase Complex (100x)
5. Sample Diluent
6. Antibody Diluent
7. Avidin-Biotin-Peroxidase Diluent
8. Color Developing Reagent (TMB)
9. Stop Solution
10. Wash Buffer (25x)
11. Adhesive plate sealers

Benötigtes Material: Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional). Pipettes and pipette tips capable of precisely dispensing 0.5 μL through 1 mL volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500 mL graduated cylinders. Test tubes for dilution.

Anwendungsinformationen

Applikationshinweise: 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.

Probenmenge: 100 μL

Plattentyp: Pre-coated

Protokoll: ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. An antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells, a biotinylated detection antibody specific for target 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 target amount in sample captured in plate.

Testpräzision: Intra-Assay Precision (CV%): 4.1%, 4.8%, 7.3%
Inter-Assay Precision (CV%): 5.7%, 5.6%, 8.9%

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (ELISA)

Validierung #101868 (ELISA)

by: Kinderklinik, Universitätsklinikum, TU Dresden

No.: #101868

Datum: 16.05.2018

Antigen: IL1B

Chargennummer: 1141380914

Validierte Anwendung: ELISA

Positivkontrolle: Plasma from LPS stimulated human whole blood; Recombinant human IL-1B with increasing concentrations of canakinumab.

Negativkontrolle: Performed following the instructions provided by the manufacturer.

Bewertung:

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.

Validierungsbilder

A. Human anticoagulated whole blood was stimulated with LPS or LPS and ATP. Subsequently, IL-1B plasma concentration was analyzed using ABIN411293. B. Recombinant human IL-1B was incubated for 2h with increasing doses of canakinumab. Subsequently, IL-1b concentration was analyzed using ABIN411293.

Protokoll

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.

Anmerkungen:

• 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.

Handhabung

Handhabung: Avoid multiple freeze-thaw cycles.

Lagerung: -20 °C,4 °C

Informationen zur Lagerung: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Haltbarkeit: 12 months

Referenzen für IL-1 beta ELISA Kit (ABIN411293)

Gao, Huang, Zhao, Hu, Li, Guo, Zhao, Lu: "LL202 protects against dextran sulfate sodium-induced experimental colitis in mice by inhibiting MAPK/AP-1 signaling." in: Oncotarget, Vol. 7, Issue 39, pp. 63981-63994, (2018) (PubMed).

Sun, Chen, Wang, Wan, Zhang, Zhang, Lin, Zhang: "Salusin-β Is Involved in Diabetes Mellitus-Induced Endothelial Dysfunction via Degradation of Peroxisome Proliferator-Activated Receptor Gamma." in: Oxidative medicine and cellular longevity, Vol. 2017, pp. 6905217, (2018) (PubMed).

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." in: 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." in: Experimental and therapeutic medicine, Vol. 15, Issue 3, pp. 2429-2435, (2018) (PubMed).

Wang, Xie, Yang, Zhang, Wang, Wu, Shen, Xie: "Sulfated Cyclocarya paliurus polysaccharides markedly attenuates inflammation and oxidative damage in lipopolysaccharide-treated macrophage cells and mice." in: Scientific reports, Vol. 7, pp. 40402, (2017) (PubMed).

Jian, Dai, Hu, Yao, Zheng, Zhu: "ox-LDL increases microRNA-29a transcription through upregulating YY1 and STAT1 in macrophages." in: 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." in: 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." in: 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." in: International journal of molecular sciences, Vol. 18, Issue 4, (2017) (PubMed).

Ballerini, Diomede, Petragnani, Cicchitti, Merciaro, Cavalcanti, Trubiani: "Conditioned medium from relapsing-remitting multiple sclerosis patients reduces the expression and release of inflammatory cytokines induced by LPS-gingivalis in THP-1 and MO3.13 cell lines." in: Cytokine, Vol. 96, pp. 261-272, (2017) (PubMed).

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." in: 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)." in: 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." in: Antioxidants & redox signaling, Vol. 26, Issue 1, pp. 28-43, (2017) (PubMed).

Bao, Li, Luo, Tang, Lv, Li, Zhou: "NF-κB-Regulated miR-99a Modulates Endothelial Cell Inflammation." in: Mediators of inflammation, Vol. 2016, pp. 5308170, (2017) (PubMed).

Song, Sun, Wang, Yang, Huang, Fu, Gan, Huang: "Pannexin3 inhibits TNF-α-induced inflammatory response by suppressing NF-κB signalling pathway in human dental pulp cells." in: Journal of cellular and molecular medicine, Vol. 21, Issue 3, pp. 444-455, (2017) (PubMed).

Xuanfei, Hao, Zhujun, Yanming, Jianping: "Imidazoline I2 receptor inhibitor idazoxan regulates the progression of hepatic fibrosis via Akt-Nrf2-Smad2/3 signaling pathway." in: Oncotarget, Vol. 8, Issue 13, pp. 21015-21030, (2017) (PubMed).

Qin, Xu, Qi, Guo, Wang, Xu, Qiao, Gu, Niu: "Evaluation of the C-domain of heparanase during AGE-induced macrophage inflammatory response." in: Experimental and therapeutic medicine, Vol. 14, Issue 2, pp. 1017-1022, (2017) (PubMed).

Zhou, Liu, Zhang, Tang, Li, Wang, Hu: "Oroxylin A inhibits colitis by inactivating NLRP3 inflammasome." in: Oncotarget, Vol. 8, Issue 35, pp. 58903-58917, (2017) (PubMed).

Li, Tang, Shuai, Jiang, Liu, Wang, Yao, Nie, Xie: "Mannose Receptor Mediates the Immune Response to Ganoderma atrum Polysaccharides in Macrophages." in: Journal of agricultural and food chemistry, Vol. 65, Issue 2, pp. 348-357, (2016) (PubMed).

Zhang, Li, Li, Shi, Zhou, Fu, Zhang, Yang, Fu, Lu: "Hypericin-photodynamic therapy induces human umbilical vein endothelial cell apoptosis." in: Scientific reports, Vol. 5, pp. 18398, (2016) (PubMed).

Details zu IL-1 beta

Target: IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))

Andere Bezeichnung: IL1B

Hintergrund:

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.

Gene Full Name: interleukin 1 beta

Gen-ID: 3553

UniProt: P01584

Pathways: NF-kappaB Signalweg, Interferon-gamma Pathway, TLR Signalweg, 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, Autophagie, Cancer Immune Checkpoints, Inflammasome