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    human-il-33-elisa-kit-ab223865.pdf

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RecombinantSimpleStep

Human IL-33 ELISA Kit (ab223865)

  • Datasheet
  • SDS
  • Protocol Booklet
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Other - Human IL-33 ELISA Kit (ab223865)
  • Example of human IL33 standard curve in Sample Diluent 50BP.
  • Example of human IL33 standard curve in Sample Diluent NS.
  • Example of human IL33 standard curve in 1X Cell Extraction Buffer PTR.
  • Interpolated concentrations of spike IL33 in human serum, plasma and cell culture media (RPMI + 10% FBS) samples.
  • Interpolated concentrations of native IL33 in human PMA/LPS stimulated THP-1 cell extract based on a 1,000 µg/mL extract load.
  • Comparison of IL33 in unstimulated and PMA/LPS stimulated THP-1 cell supernatants.
  • Sandwich ELISA - Human IL-33 ELISA Kit (ab223865)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 4.7 pg/ml
  • Range: 11.72 pg/ml - 750 pg/ml
  • Sample type: Cell culture extracts, Cell culture supernatant, Plasma, Serum
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human IL-33 ELISA Kit
    See all IL-33 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Spiked Serum 3 2.7%
    Inter-assay
    Sample n Mean SD CV%
    Spiked Serum 5 3.2%
  • Sample type

    Cell culture supernatant, Serum, Plasma, Cell culture extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    4.7 pg/ml
  • Range

    11.72 pg/ml - 750 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 87 83% - 95%
    Cell culture extracts 95 92% - 100%
    Cell culture media 102 96% - 108%
    Hep Plasma 102 97% - 110%
    EDTA Plasma 106 100% - 112%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human IL-33 ELISA Kit (ab223865) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of IL-33 protein in cell culture supernatant, plasma, serum, and cell culture extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human IL-33 with 4.7 pg/ml sensitivity.


    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:


            - Single-wash protocol reduces assay time to 90 minutes or less
            - High sensitivity, specificity and reproducibility from superior antibodies
            - Fully validated in biological samples
            - 96-wells plate breakable into 12 x 8 wells strips


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

  • Notes

    IL-33 (Interleukin-33, also known as IL-1F11 or NF-HEV) is a proinflammatory cytokine member of the IL-1 family. It is widely expressed and functions by binding and signaling through the IL-1RL1/ST2 receptor. IL-33 is expressed as a pro-protein; the antibodies in this kit were generated using the mature form of IL-33 (aa 109-270).  

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BI 1 x 6ml
    10X Human IL-33 Capture Antibody 1 x 600µl
    10X Human IL-33 Detector Antibody 1 x 600µl
    Human IL-33 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent 50BP 1 x 20ml
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Cytokines
    • Other
    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Other
  • Function

    Cytokine that binds to and signals through the IL1RL1/ST2 receptor which in turn activates NF-kappa-B and MAPK signaling pathways in target cells (PubMed:16286016). Involved in the maturation of Th2 cells inducing the secretion of T-helper type 2-associated cytokines. Also involved in activation of mast cells, basophils, eosinophils and natural killer cells. Acts as a chemoattractant for Th2 cells, and may function as an "alarmin", that amplifies immune responses during tissue injury (PubMed:17853410, PubMed:18836528).
    In quiescent endothelia the uncleaved form is constitutively and abundantly expressed, and acts as a chromatin-associated nuclear factor with transcriptional repressor properties, it may sequester nuclear NF-kappaB/RELA, lowering expression of its targets (PubMed:21734074). This form is rapidely lost upon angiogenic or proinflammatory activation (PubMed:18787100).
  • Tissue specificity

    Expressed at high level in high endothelial venules found in tonsils, Peyer patches and mesenteric lymph nodes. Almost undetectable in placenta.
  • Sequence similarities

    Belongs to the IL-1 family. Highly divergent.
  • Domain

    The homeodomain-like HTH domain mediates nuclear localization and heterochromatin association.
  • Post-translational
    modifications

    The full length protein can be released from cells and is able to signal via the IL1RL1/ST2 receptor. However, proteolytic processing by CSTG/cathepsin G and ELANE/neutrophil elastase produces C-terminal peptides that are more active than the unprocessed full length protein. May also be proteolytically processed by calpains (PubMed:19596270). Proteolytic cleavage mediated by apoptotic caspases including CASP3 and CASP7 results in IL33 inactivation (PubMed:19559631). In vitro proteolytic cleavage by CASP1 was reported (PubMed:16286016) but could not be confirmed in vivo (PubMed:19465481) suggesting that IL33 is probably not a direct substrate for that caspase.
  • Cellular localization

    Nucleus. Chromosome. Cytoplasmic vesicle, secretory vesicle. Secreted. Associates with heterochromatin and mitotic chromosomes (PubMed:17185418).
  • Target information above from: UniProt accession O95760 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • C9orf26
    • CHROMOSOME 9 OPEN READING FRAME 26
    • DKFZp586H0523
    • DVS27
    • DVS27 related protein
    • IL 1F11
    • IL 33
    • IL-1F11
    • IL-33
    • IL1F11
    • IL33
    • IL33_HUMAN
    • Interleukin 1 family member 11
    • Interleukin 33
    • INTERLEUKIN 33 NFHEV
    • Interleukin 33 precursor
    • Interleukin-1 family member 11
    • Interleukin-33 (109-270)
    • Interleukin33
    • NF HEV
    • NF-HEV
    • NFEHEV
    • NFHEV
    • Nuclear factor for high endothelial venules
    • Nuclear factor from high endothelial venules
    • OTTHUMP00000021041
    • RP11 575C20.2
    see all
  • Database links

    • Entrez Gene: 90865 Human
    • Omim: 608678 Human
    • SwissProt: O95760 Human
    • Unigene: 731660 Human

    Associated products

    • Alternative Versions

      • Human IL-33 Antibody Pair - BSA and Azide free (ab244022)

    Images

    • Other - Human IL-33 ELISA Kit (ab223865)
      Other - Human IL-33 ELISA Kit (ab223865)

      SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

       

    • Example of human IL33 standard curve in Sample Diluent 50BP.
      Example of human IL33 standard curve in Sample Diluent 50BP.

      Background-subtracted data values (mean +/- SD) are graphed.

    • Example of human IL33 standard curve in Sample Diluent NS.
      Example of human IL33 standard curve in Sample Diluent NS.

      Background-subtracted data values (mean +/- SD) are graphed.

    • Example of human IL33 standard curve in 1X Cell Extraction Buffer PTR.
      Example of human IL33 standard curve in 1X Cell Extraction Buffer PTR.

      Background-subtracted data values (mean +/- SD) are graphed.

    • Interpolated concentrations of spike IL33 in human serum, plasma and cell culture media (RPMI + 10% FBS) samples.
      Interpolated concentrations of spike IL33 in human serum, plasma and cell culture media (RPMI + 10% FBS) samples.

      The concentrations of IL33 were measured in duplicates, interpolated from the IL33 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum neat, plasma (heparin) neat, plasma (EDTA) neat and RPMI + 10% FBS neat. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

    • Interpolated concentrations of native IL33 in human PMA/LPS stimulated THP-1 cell extract based on a 1,000 µg/mL extract load.
      Interpolated concentrations of native IL33 in human PMA/LPS stimulated THP-1 cell extract based on a 1,000 µg/mL extract load.

      The concentrations of IL33 were measured in duplicate and interpolated from the IL33 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL33 concentration was determined to be 351.5 pg/mL in PMA/LPS stimulated THP-1 cell extract and undetectable in unstimulated THP-1 cell extract (not shown).

    • Comparison of IL33 in unstimulated and PMA/LPS stimulated THP-1 cell supernatants.
      Comparison of IL33 in unstimulated and PMA/LPS stimulated THP-1 cell supernatants.

      THP-1 cells were cultured for 24 hours in the presence or absence of 50 ng/mL PMA and 10 µg/mL LPS. The concentrations of IL33 were measured in neat supernatant samples in quadruplicates and interpolated from the IL33 standard curve. The interpolated values are plotted (mean +/- SD, n=3). The mean IL33 concentration was determined to be 15.95 pg/mL in neat PMA/LPS stimulated THP-1 cell culture supernatant and undetectable in neat unstimulated THP-1 cell culture supernatant and RPMI media (not shown).

    • Sandwich ELISA - Human IL-33 ELISA Kit (ab223865)
      Sandwich ELISA - Human IL-33 ELISA Kit (ab223865)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (4)

    Publishing research using ab223865? Please let us know so that we can cite the reference in this datasheet.

    ab223865 has been referenced in 4 publications.

    • Ishiguro N  et al. Activated M2 Macrophages Contribute to the Pathogenesis of IgG4-Related Disease via Toll-like Receptor 7/Interleukin-33 Signaling. Arthritis Rheumatol 72:166-178 (2020). PubMed: 31339007
    • Nian JB  et al. Epithelial cells expressed IL-33 to promote degranulation of mast cells through inhibition on ST2/PI3K/mTOR-mediated autophagy in allergic rhinitis. Cell Cycle 19:1132-1142 (2020). PubMed: 32298206
    • Li L  et al. Bone changes and curative effect of infliximab in patients with ankylosing spondylitis. J Musculoskelet Neuronal Interact 20:437-443 (2020). PubMed: 32877981
    • Zhang L  et al. Metformin Can Alleviate the Symptom of Patient with Diabetic Nephropathy Through Reducing the Serum Level of Hcy and IL-33. Open Med (Wars) 14:625-628 (2019). PubMed: 31535033

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