Human IL-33 ELISA Kit (ab223865)
- Datasheet
- References
- Protocols
Overview
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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% Heparin Plasma 102 97% - 110% EDTA Plasma 106 100% - 112% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Does not react with: Mouse -
Product overview
IL-33 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-33 protein in human serum, plasma, cell culture supernatants, and cell 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 substrate 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 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 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 (aa 109-270).
Sensitivity:
Samples diluted in Sample Diluent 50BP - 4.7 pg/mL
Samples diluted in Sample Diluent NS - 5.1 pg/mL
Samples diluted in 1X Cell Extraction Buffer PTR - 4.9 pg/mL
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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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 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
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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
modificationsThe 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
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Alternative names
- C9orf26
- CHROMOSOME 9 OPEN READING FRAME 26
- DKFZp586H0523
see all -
Database links
- Entrez Gene: 90865 Human
- Omim: 608678 Human
- SwissProt: O95760 Human
- Unigene: 731660 Human
Associated products
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Alternative Versions
Applications
Our Abpromise guarantee covers the use of ab223865 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
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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.
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Example of human IL33 standard curve in Sample Diluent 50BP.Background-subtracted data values (mean +/- SD) are graphed.
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Example of human IL33 standard curve in Sample Diluent NS.Background-subtracted data values (mean +/- SD) are graphed.
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Example of human IL33 standard curve in 1X Cell Extraction Buffer PTR.Background-subtracted data values (mean +/- SD) are graphed.
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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).
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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).
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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).
Protocols
References
ab223865 has not yet been referenced specifically in any publications.
