Human IL-1 beta/IL-1F2 Quantikine HS ELISA Kit

  
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    6.5 hours
  • Sample Type & Volume Required Per Well
    Serum (150 uL), EDTA Plasma (150 uL), Heparin Plasma (150 uL)
  • Sensitivity
    0.14 pg/mL
  • Assay Range
    0.1 - 8 pg/mL (Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant human IL-1 beta
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine HS Human IL-1 beta Immunoassay is a 6.5 hour solid phase ELISA designed to measure human IL-1 beta levels in serum and plasma. It contains E. coli-expressed recombinant human IL-1 beta and antibodies raised against the recombinant factor and has been shown to accurately quantitate recombinant human IL-1 beta. Results obtained using natural IL-1 beta showed linear curves that were parallel to the standard curves obtained using the Quantikine HS kit standards. These results indicate that this kit can be used to determine relative mass values for natural IL-1 beta. Reports indicate that ELISA kits calibrated using mature IL-1 beta as a standard will detect, but considerably underestimate, the unprocessed IL-1 beta precursor present in samples. In biological samples other than cell lysates, the precursor form of IL-1 beta is usually not the predominant form of IL-1 beta present and, additionally, is not biologically active. Therefore, results obtained using this kit should provide a useful measure of the levels of active IL-1 beta present in serum and plasma.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean0.2841.7253.2690.3061.6553.195
Standard Deviation0.0290.0960.140.0320.1210.29
CV%10.25.64.310.47.39.1

Serum, EDTA Plasma, Heparin Plasma, Saliva
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean0.3151.32.520.271.362.6
Standard Deviation0.0140.050.060.0290.1010.163
CV%4.43.92.410.77.46.3

Recovery

The recovery of IL-1 beta was determined by spiking to levels throughout the range of the assay in various matrices.

Sample Type Average % Recovery Range %
EDTA Plasma (n=4) 101 87-112
EDTA Plasma (n=4) 101 87-112
Heparin Plasma (n=4) 103 85-117
Heparin Plasma (n=4) 103 85-117
Serum (n=4) 103 95-111
Serum (n=4) 103 95-111
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of IL-1 beta and serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human IL-1 beta/IL-1F2 Quantikine HS ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-1 beta/IL-1F2
IL-1 beta (Interleukin-1 beta) is produced in response to inflammatory agents, infections, or microbial endotoxins. It plays a central role in immune and inflammatory responses, bone remodeling, fever, carbohydrate metabolism, and GH/IGF-I physiology. IL-1 beta dysregulation is implicated in many pathological conditions including sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myelogenous leukemia, insulin-dependent diabetes mellitus, atherosclerosis, neuronal injury, and aging-related diseases. Intracellular cleavage of the IL-1 beta precursor by Caspase-1/ICE is a key step in the inflammatory response. IL-1 beta binds to IL-1 RI and IL-1 RII. The IL-1 receptor accessory protein (IL-1 RAcP) associates with IL-1 RI and is required for IL-1 RI signal transduction, and IL-1ra is a secreted molecule that functions as a competitive inhibitor of IL-1 bioactivity.
    • Long Name
      Interleukin 1 beta
    • Entrez Gene IDs
      3553 (Human); 16176 (Mouse); 24494 (Rat); 397122 (Porcine); 403974 (Canine); 100034237 (Equine); 100135556 (Guinea Pig);
    • Alternate Names
      catabolin; IL-1 beta; IL-1; IL1B; IL-1B; IL1-BETA; IL-1F2; IL1F2IL-1 beta; interleukin 1, beta; interleukin-1 beta; preinterleukin 1 beta; pro-interleukin-1-beta;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 150 µL Standard, Control, or Sample
    6.   Add 150 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 5 times for a total of 6 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 6 times.

    11. 50 µL Substrate Solution
    12.   Add 50 µL Substrate Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour on the shaker. Do not wash the plate.

    13. 50 µL Amplifier Solution
    14.   Add 50 µL Amplifier Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes on the shaker.

    15. 50 µL Stop Solution
    16. Add 50 µL of Stop Solution to each well. Read at 490 nm within 30 minutes. Set wavelength correction to 650 nm or 690 nm.

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