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Mouse/Rat IL-33

Mouse/Rat IL-33 Quantikine ELISA Kit Summary

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 uL), Serum (50   uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)

Sensitivity

14.3 pg/mL

Assay Range

31.2 - 2,000 pg/mL (Cell Culture Supernates,   Serum, EDTA Plasma, Heparin Plasma)

Specificity

Natural and recombinant mouse and rat IL-33

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 Mouse/Rat IL-33 immunoassay is a 4.5 hour solid-phase ELISA designed to measure IL-33 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse mature IL-33 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate both recombinant mouse and rat IL-33. Results obtained using natural IL-33 showed dose response curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural IL-33.

Preparation and Storage

Shipping

The product is shipped at ambient   temperature. Upon receipt, store it immediately at the temperature   recommended below.

Storage

Store the unopened product at 2 - 8 °C. Do   not use past expiration date.

Background: IL-33

IL-33 (Interleukin-33) is released from physically damaged or necrotic cells. It triggers Th2-biased immune cell activation at sites of inflammation as well as regulatory T cell and M2 macrophage expansion. It additionally induces angiogenesis, promotes tumor cell migration and invasion, limits cardiac myocyte hypertrophy, and limits the development of atherosclerotic plaques. IL-33 signals through a receptor complex composed of ST2 and IL-1 RAcP. IL-1 RAcP also associates with IL-1 RI, IL-1 RII, IL-1 R6, and SCF R/c-kit. Soluble isoforms of ST2 and IL-1 RAcP function as decoy receptors that regulate IL-33 bioactivity.

Long Name:

Interleukin 33

Entrez Gene IDs:

90865 (Human); 77125 (Mouse)

Alternate Names:

C9orf26; C9orf26chromosome 9 open reading   frame 26 (NF-HEV); DKFZp586H0523; DVS27; DVS27-related protein; IL1F11;   IL-1F11; IL33; IL-33; interleukin 33; Interleukin-1 family member 11;   interleukin-33; NFHEV; NF-HEV; NF-HEVNFEHEV; Nuclear factor from high   endothelial venules; RP11-575C20.2

Assay Procedure

Refer to the product datasheet for the 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.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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