Overview

  • Product name

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

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 2.5%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 3%
  • Sample type

    Cell culture supernatant, Serum, Tissue Extracts, Heparin Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    6 pg/ml
  • Range

    31.3 pg/ml - 3000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 117 106% - 123%
    Serum 99 98% - 101%
    Tissue Extracts 87 85% - 92%
    Cell culture media 117 110% - 121%
    Heparin Plasma 82 81% - 83%
    Citrate Plasma 103 101% - 106%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Rat
    Does not react with: Human
  • Product overview

    IL-33 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-33 protein in ratserum, plasma, tissue culture supernatant, and tissue extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB Development Solution is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    IL-33 (Interleukin-33), also known as NF-HEV, is a proinflammatory cytokine member of the IL-1 family. It is widely expressed and functions by binding and signaling through the IL1RL1/ST2 receptor. IL-33 is expressed as a pro-protein; the antibodies in this kit were generated using the mature form of IL-33.


    Sensitivity:


    Samples in Sample Diluent NS: 8.5 pg/mL.


    Samples in 1X Cell Extraction Buffer PTR: 7.0 pg/mL.


    Samples in Sample Diluent 50BP: 6.0 pg/mL.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • 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 Rat IL-33 Capture Antibody 1 x 600µl
    10X Rat IL-33 Detector Antibody 1 x 600µl
    Rat IL-33 Lyophilized Recombinant Protein 2 vials
    Antibody Diluent 5BR 1 x 6ml
    10X Wash Buffer PT (ab206977) 1 x 20ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    50X Cell Extraction Enhancer Solution 1 x 1ml
    TMB Development Solution 1 x 12ml
    Stop Solution 1 x 12ml
    Sample Diluent NS (ab193972) 1 x 50ml
    Sample Diluent 50BP 1 x 20ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Plate Seals 1 unit
  • Research areas

  • 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).
  • 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

Associated products

Applications

Our Abpromise guarantee covers the use of ab236714 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • 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.

     

  • Standard Curve comparison between Rat IL-33 SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity.

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

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

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

  • Interpolated concentrations of native IL-33 in stimulated (LPS) and unstimulated (mock) 6 days rat lung tissue culture supernatant samples and stimulated and unstimulated 18 hours rat spleen tissue supernatant samples. The concentrations of IL-33 were measured in duplicates, interpolated from the IL-33 standard curves and corrected for sample dilution. Undiluted samples are as follows: lung (LPS) neat, lung (mock) neat, spleen (LPS) neat, and spleen (mock) neat. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-33 concentration was determined to be 931.1 pg/mL in LPS stimulated lung tissue supernatant, 940.4 pg/mL in unstimulated rat lung tissue supernatant, 1,503 pg/mL in LPS stimulated spleen tissue supernatant, and 1,185 pg/mL in unstimulated spleen tissue supernatant.

  • The concentrations of IL-33 were measured in duplicate and interpolated from the IL-33 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-33 concentration was determined to be 464.4 pg/mL in kidney tissue extract.

  • Lung tissue was cultured for 6 days in the presence or absence (mock) of 5 µg/mL LPS. Spleen tissue was cultured for 18 hours in the presence or absence (mock) of 1 µg/mL LPS. The concentrations of IL-33 were measured in three different dilutions of the supernatant samples in duplicates and interpolated from the IL-33 standard curve. The interpolated values are plotted (mean +/- SD, n=3). The mean IL-33 concentration was determined to be 965.8 pg/mL in LPS stimulated lung tissue culture supernatant, 940.4 in unstimulated lung tissue culture supernatant, 1,463 pg/mL in LPS stimulated spleen tissue culture supernatant, 1,165 pg/mL in unstimulated spleen tissue culture supernatant, and undetectable in RPMI1640 media containing 10% FBS (not shown).

Protocols

References

ab236714 has not yet been referenced specifically in any publications.

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