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IL-4 ELISA Kit

IL4 Reactivity: Mouse Colorimetric Sandwich ELISA Cell Lysate, Tissue Lysate
Catalog No. ABIN625148
  • Target See all IL-4 (IL4) ELISA Kits
    IL-4 (IL4) (Interleukin 4 (IL4))
    Reactivity
    • 13
    • 8
    • 6
    • 5
    • 4
    • 4
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    Mouse
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Application
    ELISA
    Purpose
    Mouse IL-4 ELISA Kit for cell and tissue lysate samples.
    Sample Type
    Tissue Lysate, Cell Lysate
    Analytical Method
    Quantitative
    Specificity
    The antibody pair provided in this kit recognizes mouse IL-4.
    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
    • Cell lysate buffer
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  • 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: Tissue lysate and cell lysate sample should be diluted at least 5-fold with Assay Diluent.
      3. Sample Diluent Buffer (Item D) and Assay Diluent (Item E) 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 1x Sample Diluent Buffer (Item D should be diluted 5-fold with deionized or distilled water before use) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 4 µL IL-4 standard from the vial of Item C, into a tube with 996 µL 1x Assay Diluent to prepare a 200 pg/mL stock standard solution. Pipette 450 µL 1x Sample Diluent Buffer into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Sample Diluent Buffer serves as the zero standard (0 pg/mL). 300 µL 300myl 300 µL 300 µL 300 µL 300 µL 4 µL standard +996 µL 200 80 32 12.8 5.12 2.05 0.819 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 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 100-fold with 1x Assay Diluent and used in step 4 of Part VI Assay Procedure.
      7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) before use. HRP-Streptavidin concentrate should be diluted 600-fold with 1x Assay Diluent . For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 20 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent to prepare a 600-fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.
      8. Cell lysate buffer is diluted to 2-fold with deionized or distilled water (for cell lysate and tissue lysate).
    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 Mouse IL-4 concentration (pg/mL) 0.1 1 10 100 1000 O D =4 50 n m 0.1 1 10
    Sensitivity: The minimum detectable dose of IL-4 is typically less than 1 pg/mL.
    Recovery: Recovery was determined by spiking various levels of mouse IL-4 into tissue lysate and cell lysate. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Tissue lysate 91.57 82-103 Cell lysate 93.62 84-104
    Linearity: Sample Type Tissue Cell Lysate lysate 1:2 Average % of 92 93 Expected Range ( %) 82-105 83-104 1:4 Average % of 94 92 Expected Range ( %) 83-104 82-103
    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
  • Wu, You, Ma, Li, Yuan, Li, Ye, Liu, Yao, Chen, Lai, Yang: "Role of transient receptor potential ion channels and evoked levels of neuropeptides in a formaldehyde-induced model of asthma in BALB/c mice." in: PLoS ONE, Vol. 8, Issue 5, pp. e62827, (2013) (PubMed).

    Herrmann, Andreeff, Gruss, Brach, Lübbert, Mertelsmann: "Interleukin-4 inhibits growth of multiple myelomas by suppressing interleukin-6 expression." in: Blood, Vol. 78, Issue 8, pp. 2070-4, (1991) (PubMed).

    Karray, DeFrance, Merle-Béral, Banchereau, Debré, Galanaud: "Interleukin 4 counteracts the interleukin 2-induced proliferation of monoclonal B cells." in: The Journal of experimental medicine, Vol. 168, Issue 1, pp. 85-94, (1988) (PubMed).

    Paul, Ohara: "B-cell stimulatory factor-1/interleukin 4." in: Annual review of immunology, Vol. 5, pp. 429-59, (1987) (PubMed).

  • Target See all IL-4 (IL4) ELISA Kits
    IL-4 (IL4) (Interleukin 4 (IL4))
    Alternative Name
    IL-4 (IL4 Products)
    Synonyms
    IL4 ELISA Kit, IL-4 ELISA Kit, BCGF-1 ELISA Kit, BCGF1 ELISA Kit, BSF-1 ELISA Kit, BSF1 ELISA Kit, Il-4 ELISA Kit, Il4e12 ELISA Kit, IL2 ELISA Kit, interleukin 4 ELISA Kit, IL4 ELISA Kit, Il4 ELISA Kit
    Background
    Mouse Interleukin-4 (IL-4) is a 20 kDa single chain glycoprotein that is synthesized as a 140 amino acid. IL-4 enhances expression of class II MHC antigens on B-cells. It can promote their capacity to respond to other B-cell stimuli and to present antigens for T-cells. IL-4 inhibits cell activation of NK-cells induced by IL-2. IL-4 stimulates the proliferation of thymocytes with the marker spectrum CD4 (-) CD8 (-), CD4 (+) CD8 (-), CD4 (-) CD8(+). The Mouse IL-4 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse IL-4 in cell lysate and tissue lysate. This assay employs an antibody specific for mouse IL-4 coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-4 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse IL-4 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 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
    16189
    UniProt
    P07750
    Pathways
    JAK-STAT Signaling, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Proton Transport, Activated T Cell Proliferation
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