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IL-2 ELISA Kit

IL2 Reactivity: Mouse Colorimetric Sandwich ELISA 1-40 pg/mL Cell Culture Supernatant, Plasma, Serum
Catalog No. ABIN1979749
  • Target See all IL-2 (IL2) ELISA Kits
    IL-2 (IL2) (Interleukin 2 (IL2))
    Reactivity
    • 13
    • 12
    • 7
    • 5
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Mouse
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Detection Range
    1-40 pg/mL
    Minimum Detection Limit
    1 pg/mL
    Application
    ELISA
    Purpose
    Mouse IL-2 ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Sample Type
    Serum, Plasma, Cell Culture Supernatant
    Analytical Method
    Quantitative
    Specificity
    This ELISA kit shows no cross-reactivity with any of the cytokines tested: Mouse CD30, L CD30, T CD40, CRG-2, CTACK, CXCL16, Eotaxin , Eotaxin-2, Fas Ligand, Fractalkine, GCSF, GM-CFS, IFN-gamma, IGFBP-3, IGFBP-5, IGFBP-6, IL-1 alpha, IL-1 beta, IL-3, IL-3 Rb, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40/p70, IL-12 p70, IL-13, IL-17, KC, Leptin R, LEPTIN(OB), LIX, L-Selectin, Lymphotactin, MCP-1, MCP-5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, MIP-2, MIP-3 beta, MIP-3 alpha, PF-4, P-Selectin, RANTES, SCF, SDF-1 alpha, TARC, TCA-3, TECK, TIMP-1, TNF-alpha, TNF RI, TNF RII, TPO, VCAM-1, VEGF.
    Sensitivity
    < 1 pg/mL
    Characteristics
    • Strip plates and additional reagents allow for use in multiple experiments
    • Quantitative protein detection
    • Establishes normal range
    • The best products for confirmation of antibody array data
    Components
    • Pre-Coated 96-well Strip Microplate
    • Wash Buffer
    • Stop Solution
    • Assay Diluent(s)
    • Lyophilized Standard
    • Biotinylated Detection Antibody
    • Streptavidin-Conjugated HRP
    • TMB One-Step Substrate
    Material not included
    • Distilled or deionized water
    • Precision pipettes to deliver 2 μL to 1 μL volumes
    • Adjustable 1-25 μL pipettes for reagent preparation
    • 100 μL and 1 liter graduated cylinders
    • Tubes to prepare standard and sample dilutions
    • Absorbent paper
    • Microplate reader capable of measuring absorbance at 450nm
    • Log-log graph paper or computer and software for ELISA data analysis
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  • Application Notes
    Recommended Dilution for serum and plasma samples2 fold
    Sample Volume
    100 μL
    Plate
    Pre-coated
    Protocol
    1. Prepare all reagents, samples and standards as instructed in the manual.
    2. Add 100 μL of standard or sample to each well.
    3. Incubate 2.5 h at RT or O/N at 4 °C.
    4. Add 100 μL of prepared biotin antibody to each well.
    5. Incubate 1 h at RT.
    6. Add 100 μL of prepared Streptavidin solution to each well.
    7. Incubate 45 min at RT.
    8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 μL of Stop Solution to each well.
    11. Read at 450 nm immediately.
    Reagent Preparation
    1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
      2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants. Suggested dilution for normal serum/plasma: 2 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
      3. Assay Diluent B should be diluted 5-fold with deionized or distilled water before use.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium and urine) into Item C vial to prepare a 12 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 2 µL IL-2 standard (12 ng/mL) from the vial of Item C, into a tube with 598 µL Assay Diluent A or 1x Assay Diluent B to prepare 40 pg/mL stock standard solution. Pipette 300 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 300 µL 2 µL standard +598 µL 300myl 300 µL 300 µL 300 µL 300 µL 40 20 10 5 2.5 1.25 0.625 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
      5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
      6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
      7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 120-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 100 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a final 120 fold diluted HRP-Streptavidin solution. Mix well.
    Assay Procedure
    1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
      2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
      3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
      4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
      5. Discard the solution. Repeat the wash as in step
      6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
      7. Discard the solution. Repeat the wash as in step
      8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
      9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
    Calculation of Results

    Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
    Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent A Mouse IL-2 concentration (pg/mL) 0.1 1 10 100 O D =4 50 n m 0.1 1 10 Assay Diluent B Mouse IL-2 concentration (pg/mL) 0.1 1 10 100 O D =4 50 n m 0.1 1 10
    Sensitivity: The minimum detectable dose of IL-2 is typically less than 1 pg/mL.
    Recovery: Recovery was determined by spiking various levels of mouse IL-2 into mouse serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 97.56 85-104 Plasma 98.34 86-106 Cell culture media 100.34 89-110
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 98 96 98 Range ( %) 90-105 89-107 88-106 1:4 Average % of Expected 98 95 96 Range ( %) 90-107 89-106 91-108
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    Assay Precision
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Restrictions
    For Research Use only
  • Handling Advice
    Avoid repeated freeze-thaw cycles.
    Storage
    -20 °C
    Storage Comment
    The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
    Expiry Date
    6 months
  • Ahmad, Attia, Bakheet, Zoheir, Ansari, Korashy, Abdel-Hamied, Ashour, Abd-Allah et al.: "Naringin attenuates the development of carrageenan-induced acute lung inflammation through inhibition of NF-?b, STAT3 and pro-inflammatory mediators and enhancement of I?B? and anti-inflammatory ..." in: Inflammation, Vol. 38, Issue 2, pp. 846-57, (2015) (PubMed).

    Dey, Chaudhuri: "Immunomodulatory activity of Nerium indicum through inhibition of nitric oxide and cyclooxygenase activity and modulation of TH1/T H2 cytokine balance in murine splenic lymphocytes." in: Cytotechnology, (2015) (PubMed).

    Li, Li, Deng, Zhao, Liu, Wang et al.: "Prime-boost vaccination with Bacillus Calmette Guerin and a recombinant adenovirus co-expressing CFP10, ESAT6, Ag85A and Ag85B of Mycobacterium tuberculosis induces robust antigen-specific immune ..." in: Molecular medicine reports, Vol. 12, Issue 2, pp. 3073-80, (2015) (PubMed).

    Gu, Xie, Wang, Peng, Lai, Yang: "Immune response induced by candidate Sarcoptes scabiei var. cuniculi DNA vaccine encoding paramyosin in mice." in: Experimental & applied acarology, Vol. 63, Issue 3, pp. 401-12, (2014) (PubMed).

    Tian, Li, Yang, Peng, Zhu, He, Chen: "In vivo immunotoxicity of SiO2@(Y0.5Gd0.45Eu0.05)2O3 as dual-modality nanoprobes." in: International journal of molecular sciences, Vol. 15, Issue 8, pp. 13649-62, (2014) (PubMed).

    Tian, Yang, Yang, Peng, Chen, Zhu, He, Li, Chen: "Toxicity evaluation of Gd2O3@SiO2 nanoparticles prepared by laser ablation in liquid as MRI contrast agents in vivo." in: International journal of nanomedicine, Vol. 9, pp. 4043-53, (2014) (PubMed).

    Liu, Tian, Luo, Yang, Xiao, Shao, Chen, Yang, Chen, Li: "Sub-10 nm monoclinic Gd2O3:Eu3+ nanoparticles as dual-modal nanoprobes for magnetic resonance and fluorescence imaging." in: Langmuir : the ACS journal of surfaces and colloids, Vol. 30, Issue 43, pp. 13005-13, (2014) (PubMed).

    Rodrigues, Figueiredo, Padrela, Cadete, Tiago, Matos, Gomes de Azevedo, Florindo, Gonçalves, Almeida: "Development of a novel mucosal vaccine against strangles by supercritical enhanced atomization spray-drying of Streptococcus equi extracts and evaluation in a mouse model." in: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft für Pharmazeutische Verfahrenstechnik e.V, Vol. 82, Issue 2, pp. 392-400, (2012) (PubMed).

    Bizargity, Liu, Wang, Hancock, Visner: "Inhibitory effects of pirfenidone on dendritic cells and lung allograft rejection." in: Transplantation, Vol. 94, Issue 2, pp. 114-22, (2012) (PubMed).

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  • Target See all IL-2 (IL2) ELISA Kits
    IL-2 (IL2) (Interleukin 2 (IL2))
    Alternative Name
    IL-2 (IL2 Products)
    Synonyms
    IL2 ELISA Kit, IL-2 ELISA Kit, il2 ELISA Kit, TCGF ELISA Kit, lymphokine ELISA Kit, Il-2 ELISA Kit, POIL2 ELISA Kit, interleukin 2 ELISA Kit, interleukin-15 ELISA Kit, IL2 ELISA Kit, il2 ELISA Kit, LOC100303688 ELISA Kit, Il2 ELISA Kit
    Background
    IL-2 is a protein of 133 amino acids (15.4 kDa). It is produced mainly by T-cells expressing the surface antigen CD4 following cell activation by mitogens or allogens under physiological conditions. IL-2 displays significant anti-tumor activity for a variety of tumor cell types since it supports the proliferation and clonal expansion of T-cells that specifically attack certain tumor types. Human IL-2 is active on mouse cells, mouse IL-2 is species-specific and is inactive on human cells. At the amino acid sequence level, there is approximately 60% similarity between mouse and human IL-2. The Mouse IL-2 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse IL-2 in serum, plasma and cell culture supernatants. This assay employs an antibody specific for mouse IL-2 coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse IL-2 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IL-2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
    Gene ID
    16183
    UniProt
    P04351
    Pathways
    JAK-STAT Signaling, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Activated T Cell Proliferation
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