IL-1 beta ELISA 试剂盒
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Quick Overview for IL-1 beta ELISA 试剂盒 (ABIN411293)
抗原
See all IL-1 beta (IL1B) ELISA试剂盒抗原表位
适用
检测方法
实验类型
检测范围
应用范围
样品类型
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最低检测浓度
- 3.9 pg/mL
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原理
- Sandwich Human IL-1 Beta/IL-1F2/IL1B ELISA Kit to quantitate Human IL1B in cell culture supernatants, serum and plasma (heparin, EDTA, citrate).
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Analytical Method
- Quantitative
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特异性
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Expression system for standard: E.coli
Immunogen sequence: A117-S269Capture antibody and Detection antibody: a monoclonal antibody from mouse, a biotinylated monoclonal antibody from mouse
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交叉反应 (详细)
- There is no detectable cross-reactivity with other relevant proteins.
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灵敏度
- < 0.15 pg/mL
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组件
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- Pre-coated 96-well strip microplate
- Standard
- Biotinylated antibody (100x)
- Avidin-Biotin-Peroxidase Complex (100x)
- Sample Diluent
- Antibody Diluent
- Avidin-Biotin-Peroxidase Diluent
- Color Developing Reagent (TMB)
- Stop Solution
- Wash Buffer (25x)
- Adhesive plate sealers
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试剂未包括
- 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.
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应用备注
- 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.
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样本量
- 100 μL
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板类型
- Pre-coated
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实验流程
- 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.
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实验精密度
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Intra-Assay Precision (CV%): 4.1%, 4.8%, 7.3%
Inter-Assay Precision (CV%): 5.7%, 5.6%, 8.9% -
限制
- 仅限研究用
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- by
- Kinderklinik, Universitätsklinikum, TU Dresden
- No.
- #101868
- 日期
- 2018.05.16
- 抗原
- IL1B
- Lot Number
- 1141380914
- Method validated
- ELISA
- Positive Control
- Plasma from LPS stimulated human whole blood; Recombinant human IL-1B with increasing concentrations of canakinumab.
- Negative Control
- Performed following the instructions provided by the manufacturer.
- Notes
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.
- Primary Antibody
- Secondary Antibody
- 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.
- Experimental 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.
生效 #101868 (ELISA)
Validation ImagesFull Methods -
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注意事项
- Avoid multiple freeze-thaw cycles.
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储存条件
- -20 °C,4 °C
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储存方法
- 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.)
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有效期
- 12 months
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- IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))
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别名
- IL1B
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背景
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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
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基因ID
- 3553
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UniProt
- P01584
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途径
- NF-kappaB Signaling, Interferon-gamma Pathway, TLR signaling, 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
抗原 See all IL-1 beta (IL1B) ELISA试剂盒
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(1 validation)