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IL-13 ELISA Kit

IL13 Reactivity: Human Colorimetric Sandwich ELISA 0.15-40 pg/mL Cell Culture Supernatant, Plasma, Serum
Catalog No. ABIN625011
  • Target See all IL-13 (IL13) ELISA Kits
    IL-13 (IL13) (Interleukin 13 (IL13))
    Reactivity
    • 12
    • 6
    • 3
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    Human
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Detection Range
    0.15-40 pg/mL
    Minimum Detection Limit
    0.15 pg/mL
    Application
    ELISA
    Purpose
    Human IL-13 ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Sample Type
    Plasma, Cell Culture Supernatant, Serum
    Analytical Method
    Quantitative
    Specificity
    This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    Sensitivity
    < 0.15 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 and urine. 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.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 432 µL of Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium) into Item C vial to prepare a 20 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 2 µL of IL-13 standard from the vial of Item C, into a tube with 998 µL of Assay Diluent A or 1x Assay Diluent B to prepare a 40 pg/mL stock standard solution. Pipette 300 µL of 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). 200 µL 200myl 200 µL 200 µL 200 µL 200 µL 2 µL standard + 998 µL 40 16 6.4 2.56 1.03 0.41 0.16 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 solutionv (don't store the diluted solution for next day use). 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 Human IL-13 concentration (pg/mL) 0.1 1 10 100 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B Human IL-13 concentration (pg/mL) 0.1 1 10 100 O D =4 50 n m 0.01 0.1 1 10
    Sensitivity: The minimum detectable dose of IL-13 is typically less than 0.15 pg/mL.
    Recovery: Recovery was determined by spiking various levels of human IL-13 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 96.35 84-103 Plasma 94.39 82-102 Cell culture media 100.65 88-107
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 95 94 98 Range ( %) 83-103 82-102 85-104 1:4 Average % of Expected 96 97 96 Range ( %) 85-104 85-105 83-102
    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
  • Akelma, Cizmeci, Kanburoglu, Bozkaya, Catal, Mete, Kutukoglu, Namuslu: "Elevated level of serum osteopontin in school-age children with asthma." in: Allergologia et immunopathologia, Vol. 42, Issue 4, pp. 275-81, (2014) (PubMed).

    Chen, Li, Xie, Huang, Yuan, Zeng: "IL-6, IL-10 and IL-13 are associated with pathogenesis in children with Enterovirus 71 infection." in: International journal of clinical and experimental medicine, Vol. 7, Issue 9, pp. 2718-23, (2014) (PubMed).

    Melloh, Lord, Staub, Merk, Barz et al.: "Re: Macedo LG, Wang Y, Battié MC. The sedimentation sign for differential diagnosis of lumbar spinal stenosis. [published online ahead of print November 28, 2012.] Spine. ..." in: Spine, Vol. 38, Issue 11, pp. 968, (2013) (PubMed).

    Hashad, Hamed, El Gharabawy, El Metwally, Morsi: "Interleukins 12 and 13 levels among beta-thalassaemia major patients." in: Eastern Mediterranean health journal = La revue de santé de la Méditerranée orientale = al-Majallah al-?i???yah li-sharq al-mutawassi?, Vol. 19, Issue 2, pp. 181-5, (2013) (PubMed).

    Barderas, Bartolomé, Fernandez-Aceñero, Torres, Casal: "High expression of IL-13 receptor ?2 in colorectal cancer is associated with invasion, liver metastasis, and poor prognosis." in: Cancer research, Vol. 72, Issue 11, pp. 2780-90, (2012) (PubMed).

    Nagayoshi, Tada, West, Ochiai, Watanabe, Toyotome, Tanabe, Takiguchi, Shigeta, Yasuda, Shibuya, Kamei, Tatsumi: "Inhalation of Stachybotrys chartarum evokes pulmonary arterial remodeling in mice, attenuated by Rho-kinase inhibitor." in: Mycopathologia, Vol. 172, Issue 1, pp. 5-15, (2011) (PubMed).

    Iikura, Katsunuma, Saika, Saito, Ichinohe, Ida, Saito, Matsumoto: "Peripheral blood mononuclear cells from patients with bronchial asthma show impaired innate immune responses to rhinovirus in vitro." in: International archives of allergy and immunology, Vol. 155 Suppl 1, pp. 27-33, (2011) (PubMed).

    Zurawski, de Vries: "Interleukin 13, an interleukin 4-like cytokine that acts on monocytes and B cells, but not on T cells." in: Immunology today, Vol. 15, Issue 1, pp. 19-26, (1994) (PubMed).

    de Waal Malefyt, Figdor, Huijbens, Mohan-Peterson, Bennett, Culpepper, Dang, Zurawski, de Vries: "Effects of IL-13 on phenotype, cytokine production, and cytotoxic function of human monocytes. Comparison with IL-4 and modulation by IFN-gamma or IL-10." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 151, Issue 11, pp. 6370-81, (1994) (PubMed).

    Cocks, de Waal Malefyt, Galizzi, de Vries, Aversa: "IL-13 induces proliferation and differentiation of human B cells activated by the CD40 ligand." in: International immunology, Vol. 5, Issue 6, pp. 657-63, (1993) (PubMed).

    Minty, Chalon, Derocq, Dumont, Guillemot, Kaghad, Labit, Leplatois, Liauzun, Miloux: "Interleukin-13 is a new human lymphokine regulating inflammatory and immune responses." in: Nature, Vol. 362, Issue 6417, pp. 248-50, (1993) (PubMed).

    Griffing: "Radiologic seminar LI: the posterior fat pad sign." in: Journal of the Mississippi State Medical Association, Vol. 7, Issue 7, pp. 310-1 (PubMed).

  • Target See all IL-13 (IL13) ELISA Kits
    IL-13 (IL13) (Interleukin 13 (IL13))
    Alternative Name
    IL-13 (IL13 Products)
    Synonyms
    IL13 ELISA Kit, IL-13 ELISA Kit, P600 ELISA Kit, Il-13 ELISA Kit, L13 ELISA Kit, interleukin 13 ELISA Kit, IL13 ELISA Kit, Il13 ELISA Kit
    Background
    Human IL-13 is expressed in activated T-helper cells and T-cells expressing CD8. It is not expressed in heart, brain, placenta, lung, liver, and skeletal muscle tissues. IL-13 is a pleiotropic cytokine that has important anti-inflammatory and immunoregulatory activities. It down-modulates macrophage activity, reducing the production of pro-inflammatory cytokines and chemokines in response to IFN-gamma or bacterial lypopolysaccharides. The Human IL-13 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human IL-13 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human IL-13 coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-13 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human IL-13 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-13 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
    3596
    UniProt
    P35225
    Pathways
    JAK-STAT Signaling, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Proton Transport
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