Human IL-33 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Interleukin 33 (IL-33) is a cytokine belonging to the IL-1 superfamily. The IL33 gene maps to chromosome 9p24.1 by using of genomic database analysis. Recombinant mature human IL33 bound to ST2. The induction of type 2 cytokines by IL-33 in vivo is believed to induce the severe pathological changes observed in mucosal organs following administration of IL-33. IL33, an alarmin released from necrotic cells, is necessary for potent CD8 + T cell (CTL) responses to replicating, prototypic RNA, and DNA viruses in mice. IL33 prevented the downregulation of CXCR2 and inhibition of chemotaxis induced by activation of TLR4, and found that IL33 reverses the TLR4-induced reduction of CXCR2 expression via the inhibition of expression of GRK2.

C9orf26; DVS27; DVS27 related protein; IL-1F11; IL-33; IL1F11; IL33; IL33_HUMAN; Interleukin-1 family member 11; Interleukin-33; NF-HEV; NFEHEV; Nuclear factor from high endothelial venules; RP11-575C20.2

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 30 260 484 28 282 511 Standard deviation 1.74 16.38 37.75 1.76 22.27 45.99 CV (%) 5.8% 6.3% 7.8% 6.3% 7.9% 9% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 30 260 484 28 282 511 Standard deviation 1.74 16.38 37.75 1.76 22.27 45.99 CV (%) 5.8% 6.3% 7.8% 6.3% 7.9% 9% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 30 260 484 28 282 511 Mean (pg/ml) 30 260 484 28 282 511 Standard deviation 1.74 16.38 37.75 1.76 22.27 45.99 Standard deviation 1.74 16.38 37.75 1.76 22.27 45.99 CV (%) 5.8% 6.3% 7.8% 6.3% 7.9% 9% CV (%) 5.8% 6.3% 7.8% 6.3% 7.9% 9%

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. 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. 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. This form is rapidely lost upon angiogenic or proinflammatory activation.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0929-CA Rabbit anti- human IL33 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0929-DA Biotinylated goat anti- human IL33 polyclonal antibody (Detection Antibody) 500 μl, 3.5 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0929-ST Lyophilized recombinant human IL33 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ0929-CA Rabbit anti- human IL33 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0929-DA Biotinylated goat anti- human IL33 polyclonal antibody (Detection Antibody) 500 μl, 3.5 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0929-ST Lyophilized recombinant human IL33 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ0929-CA Rabbit anti- human IL33 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0929-CA Rabbit anti- human IL33 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0929-DA Biotinylated goat anti- human IL33 polyclonal antibody (Detection Antibody) 500 μl, 3.5 μg/mL (recommended dilution 1:100) EZ0929-DA Biotinylated goat anti- human IL33 polyclonal antibody (Detection Antibody) 500 μl, 3.5 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0929-ST Lyophilized recombinant human IL33 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK0929-ST Lyophilized recombinant human IL33 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.029 0.057 0.101 0.189 0.335 0.632 1.152 2.098 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.029 0.057 0.101 0.189 0.335 0.632 1.152 2.098 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.029 0.057 0.101 0.189 0.335 0.632 1.152 2.098 O.D. 0.029 0.057 0.101 0.189 0.335 0.632 1.152 2.098

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)