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Mouse IL-2 ELISPOT Kit

For research use only.

组分(Materials Provided)

IDComponentsSize
RSP006-C01Pre-coated Anti-IL-2 Antibody Microplate1 plate
RSP006-C02Positive Stimulus60 μg
RSP006-C03Biotin-Anti-IL-2 Antibody50 μL
RSP006-C04Streptavidin-HRP50 μL
RSP006-C05Washing Buffer (10×)50 mL
RSP006-C06Dilution Buffer (1×)50 mL
RSP006-C07AEC Dilution 25 mL
RSP006-C08AEC Solution A (20×)0.8 mL
RSP006-C09AEC Solution B (20×)0.8 mL
RSP006-C10AEC Solution C (20×)0.8 mL

产品概述(Product Overview)

Interleukin-2 (IL-2) is a pleiotropic cytokine that drives T-cell growth, augments NK cytolytic activity, induces the differentiation of regulatory T cells, and mediates activation-induced cell death. IL-2 does not specify the type of Th differentiation that occurs; instead, IL-2 modulates expression of receptors for other cytokines and transcription factors, thereby either promoting or inhibiting cytokine cascades that correlate with each Th differentiation state. The Mouse IL-2 ELISpot assay is designed for the detection of mouse IL-2 secreting cells at the single cell level and can be used to quantitate the frequency of mouse IL-2 secreting cells. ELISpot assays are highly reproducible and sensitive, and can be used to measure responses with frequencies well below 1 in 100,000. ELISpot assays do not require prior in vitro expansion of T cells and they are suitable for high-throughput analysis using only small volumes of primary cells. As such, ELISpot assays are useful tools for research in vaccine development, cytokine secretion and the monitoring of various clinical trials.

应用说明(Application)

This kit is used to detect IL-2 at the cellular level.

It is for research use only.

重构方法(Reconstitution)

Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.

存储(Storage)

原理(Assay Principles)

ELISpot assays employ the quantitative sandwich enzyme-linked immunosorbent assay (ELISA) technique. The Kit consists of Pre-coated Anti-IL-2 Antibody Microplate, Positive Stimulus, Biotin-Anti-IL-2 Antibody, Streptavidin-HRP, AEC and buffers.

Your experiment will include 6 simple steps:

a) A monoclonal antibody specific for mouse IL-2 has been pre-coated onto a polyvinylidene difluoride (PVDF)-backed microplate.

b) Appropriately stimulated cells are pipetted into the wells and the microplate is placed into a humidified 37 °C CO2 incubator for a specified period of time.During this incubation period, the immobilized antibody in the immediate vicinity of the secreting cells binds secreted IL-2.

c) After washing away any cells and unbound substances, a biotinylated antibody specific for mouse IL-2 is added to the wells. Following a wash to remove any unbound biotinylated antibody, HRP conjugated to streptavidin is added.

d) Wash the plate and add the Streptavidin-HRP diluted by Dilution Buffer to the plate.

e) Wash the plate and add AEC. A reddish-brown colored precipitate forms and appears as spots at the sites of cytokine localization, with each individual spot representing an individual IL-2 secreting cell.

f) The spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.

质量管理控制体系(QMS)

  1. 质量管理体系(ISO, GMP)
  2. 质量优势
  3. 质控流程
 

典型数据-Typical Data Please refer to DS document for the assay protocol.

IL-2 TYPICAL DATA

1×10⁵ cells/well Mouse Spleen Cells were stimulated with 6.0 μg/mL Positive Stimulus and cultured at 37℃ and 5%CO2 for 16 h, 265 spots were produced. The following example data is for reference only.

 
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前沿进展

Effect and Mechanism of Vitamin D on Inflammatory Factors and Neutrophil Activity in Preterm Placenta of Rats Induced by LPS
Yang, Chen, Lv
Cell Biochem Biophys (2025)
Abstract: To investigate the impact mechanisms of vitamin D on inflammatory factors and neutrophil activity in preterm pregnant rats. 24 pregnant rats were selected as the research objects and randomly divided into control group, LPS group and LPS + VD group, with 8 rats in each group. On the second day of pregnancy, the LPS + VD group was injected intraperitoneally with 50 mg/L vitamin D30.2 mL, and the LPS group and the control group were injected with the same amount of 0.9% NaCl twice a day. On the seventh day of pregnancy, the LPS group and the LPS + VD group were injected with 0.2 mL LPS into the tail vein to establish a preterm labor model induced by infection. The control group was injected with the same amount of physiological saline into the tail vein. Placental tissues from rats in the LPS + VD group and the LPS group were collected, and the expression levels of inflammatory factors TGF-β1, TNF-α, and VDBP were detected by immunohistochemistry. At the same time, serum IL-2 concentration was measured by ELISA and radioimmunoassay, the activity of neutrophils was evaluated by flow cytometry, and the expression of Hippo-YAP signaling pathway protein was detected by Western blot. Compared with the control group, the content of TNF-α, VDBP, and TGF-β1 in placenta in LPS group were higher than that in the control group (P < 5); Compared with the LPS group, the contents of TNF-α, BP and TGF-β1 in the LPS+VD group were significantly reduced,(P < 0.05); Compared with the control group, the serum IL-2 concentration in the LPS group was significantly higher than that in the control group (P < 0.05); Compared with the LPS group, the serum IL-2 concentration in the LPS + VD group decreased significantly (P < 0.05); Compared with the control group, the neutrophil ratio and absolute neutrophil value in the LPS group were higher than those in the control group (P < 0.05); Compared with the LPS group, the neutrophil ratio and absolute value of neutrophils decreased (P < 0.05); compared with the control group, the expression levels of YAP and P-YAP protein in the LPS group increased (P < 0.05); compared with the LPS group, the expression levels of YAP and P-YAP protein in the LPS + VD group decreased (P < 0.05). Vitamin D can improve the immune status of preterm pregnant mice by inhibiting the expression of placental inflammatory factors.© 2025. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.
Serum Interleukin Levels Predict Occurrence of Acute Radiation Pneumonitis and Overall Survival in Thoracic Tumours
Zhang, Shen, Li et al
Clin Invest Med (2025) 48 (1), 29-38
Abstract: Radiation-induced lung injury (RILI) is a significant adverse effect of thoracic radiotherapy, potentially impacting patient prognosis. The risk factors for acute radiation pneumonitis (RP) have not been fully clarified. The present study evaluated the predictive value of serum interleukins (ILs) in the occurrence of RP and overall survival in patients with thoracic cancers.This single-centre retrospective observational study enrolled 435 thoracic cancer patients who underwent chest radiation therapy. Serum levels of IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-17, TNF-α, IFN-γ, IFN-α were measured by cytometric bead array before radiotherapy. The relationship between clinical characteristics, serum IL levels and the occurrence of RP were analyzed. Cox regression and Kaplan-Meier methods were also performed to investigate the prognostic role of serum IL levels in these patients.The incidence of RP in these patients was 17.01%. Elevated serum levels of IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, TNF-α, IFN-α were all associated with the occurrence of RP. High levels of IL-1β, IL-4, and IL-12p70 were correlated with more severe pneumonitis. Univariate and multivariate logistic regression analysis identified serum IL-6 level as an independent prognostic factor in patients receiving thoracic radiotherapy.Serum interleukin levels are linked to the development of acute RP in patients receiving thoracic radiotherapy. Serum IL-6 could serve as a valuable biomarker in identifying patients at high risk for RP, potentially guiding individualized therapeutic strategies and improving patient management in radiotherapy. Future research should focus on validating IL-6's role in larger cohorts and exploring its integration into clinical practice for the early prediction of RILI.
Upregulated TCF1+ Treg Cells With Stronger Function in Systemic Lupus Erythematosus Through Activation of the Wnt-β-Catenin
Zeng, Gao, Bahabayi et al
Immunology (2025)
Abstract: The role of T-cell factor 1 (TCF1) in human regulatory T cells (Treg) and its clinical significance in systemic lupus erythematosus (SLE) remain unclear. Through bioinformatics analysis and flow cytometry, the Tcf7 gene and TCF1 protein were found to be highly expressed in Treg cells. TCF1+ Treg cells exhibited increased expression of CTLA4 and LAG3 and higher IL-10 secretion than TCF1- Treg cells. Circulating TCF1+ Treg cells were elevated and displayed increased inhibitory markers in SLE patients. The Wnt-β-catenin pathway was activated in TCF1+ Treg cells in SLE patients. The addition of XAV939 impaired the function of TCF1+ Treg cells. Clinically, TCF1+ Treg cells were not only related to CRP, ESR and IL-2, but also could differentiate SLE patients from healthy controls, primary Sjögren's syndrome patients and rheumatoid arthritis patients. In conclusion, the increased TCF1+ Treg cells in SLE patients indicate a stronger suppressive function for the activated Wnt-β-catenin pathway and help screening and assisting in the diagnosis of SLE patients.© 2025 John Wiley & Sons Ltd.
Analysis of the therapeutic effect of acitretin capsules combined with cooling blood and detoxifying formula on psoriasis hemorrhagic fever and its impact on the gut microbiota of patients
Zhou, Li, Liu et al
Medicine (Baltimore) (2025) 104 (12), e41502
Abstract: This study evaluates the therapeutic effect of acitretin capsules combined with a cooling blood and detoxifying formula (CBDF) on psoriasis hemorrhagic fever and its impact on the gut microbiota (GM). Seventy patients diagnosed with psoriasis hemorrhagic fever between January 2022 and April 2023 were divided into 2 groups: an experimental group (EG) treated with acitretin capsules and the CBDF, and a control group treated with acitretin capsules alone. The study assessed immune inflammatory responses, symptom improvement, traditional Chinese medicine symptom scores, and changes in GM diversity and abundance before and after treatment. After 2 months, the EG showed significant reductions in interleukin-2 receptor levels (from 813.2 ± 201.5-402.3 ± 111.2 mg/L) and interleukin-6 (from 8.4 ± 3.1-3.8 ± 1.7 pg/mL). The quality of life index improved by 6.12 points in the EG, which was significantly better than the control group (P < .05). PASI scores decreased in both groups, with the EG showing greater improvement (PASI before treatment: 23.1 ± 4.3; after 1 month: 15.6 ± 3.8; after 2 months: 10.4 ± 3.3). The EG also showed a significant increase in GM diversity after treatment (P < .05). Acitretin capsules combined with the CBDF is effective in treating psoriasis hemorrhagic fever and positively affects the GM, offering a potential new treatment approach.Copyright © 2025 the Author(s). Published by Wolters Kluwer Health, Inc.
Showing 1-4 of 91326 papers.
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IL-2靶点信息
英文全称:Interleukin-2
中文全称:白细胞介素-2
种类:Homo sapiens
上市药物数量:1详情
临床药物数量:39详情
最高研发阶段:批准上市
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