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<strong>Figure 1 Western Blot Validation of IL-33 in A20 Cells </strong><br>Loading: 15 μg of lysates per lane.Antibodies: IL-33, 4273, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Lane A: 1 μg/mLLane B: 2 μg/mL
<strong>Figure 2 Western Blot Validation of IL-33 with Human Recombinant Protein</strong><br>Loading: 5ng (1) and 25ng (2) of IL-33 partial human recombinant protein.Antibodies: IL-33, 4273, 1 μg/mL , 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Il-33 partial human recombinant protein: 112-270aa, 17.9kD, 36kD band is the dimer.
<strong>Figure 3 Western Blot Validation of IL-33 in Human Lymph Node</strong><br>Loading: 15. μg of human lymph node lysateAntibodies: IL-33, 4273, 2 μg/mL , 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Western Blot Validation of IL-33 in Human Lung</strong><br>Loading: 15. μg of human lung lysateAntibodies: IL-33, 4273, 2 μg/mL , 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 5 Immunofluorescence Validation of IL-33 in Human Lymph Node</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed human lymph node tissue labeling IL-33 with 4273 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 6 Immunohistochemistry Validation of IL-33 in Human Lymph Node </strong><br>Immunohistochemical analysis of paraffin-embedded human lymph node tissue using anti-IL-33 antibody (4273) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 7 Induced Expression Validation of IL-33 in Human Skin Fibroblasts (Ivanov et al., 2010)</strong><br>Protein expression of IL-33 in human skin fibroblasts following alpha-irradiation were detected by anti-IL-33 antibodies. IL-33 levels were increased 1–4 h after α-irradiation.
<strong>Figure 8 Regulated Expression Validation of IL-33 in Human Skin Fibroblasts (Ivanov et al., 2010)</strong><br>Protein expression of IL-33 in human skin fibroblasts following alpha-irradiation were detected by anti-IL-33 antibodies. IL-33 levels were increased after α-irradiation, which were inhibited by treatment of PPP.
Figure 1 Western Blot Validation of IL-33 in A20 Cells
Loading: 15 μg of lysates per lane.Antibodies: IL-33, 4273, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Lane A: 1 μg/mLLane B: 2 μg/mL
Cell Treatment
Most Cited
Trial Size

IL-33 Antibody

CATALOG NUMBER: 4273

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Homology:
Predicted species reactivity based on immunogen sequence: Rat: (76.5%)
Immunogen:
Anti-IL-33 antibody (4273) was raised against a 17 amino acid synthetic peptide from near the center of human IL-33.
The immunogen is located within the 150-200 amino acids of IL-33.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
Application Note:
WB: 1-2 μg/mL; IF: 20 μg/mL; IHC-P: 2.5 μg/mL.
Antibody validated: Western Blot in human and mouse samples; Immunofluorescence and Immunohistochemistry in human samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1288 - A20 Cell Lysate
Positive Control 2:
Cat. No. 11-521 - Human Lymphoid Tissue Tissue Slide
Predicted Molecular Weight:
Predicted: 31kD
Observed: 33kD
Validation:

Recombinant Protein Validation (Figure 2) shows expression of human partial recombinant protein IL-33 detected by anti-IL-33 antibody, 4273.  Bands of 18kD (monomer) and 36kD (dimer) were observed.

Regulated Expression Validation (Figure 7&8) shows the up-regulation (Figure 7) of IL-33 detected by anti-IL-33 antibody in human skin fibroblasts after α-irradiation and this effect  was inhibited by PPP (Figure 8).

Isoforms:
Human IL-33 has 4 isoforms, including isoform 1 (270aa, 30.8kD), isoform 2 (228aa, 25.9kD), isoform 3 (228aa, 26.1kD) and isoform 4 (144aa, 16.3kD). This antibody detects only human isoform L. Mouse IL-33 has 1 isoform (266aa, 30kD). Rat IL-33 has only one isoform identified so far (264aa, 29.5kD).
Purification:
IL-33 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
IL-33 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
IL-33 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
IL33
Additional Names:
IL-33 Antibody: DVS27, IL1F11, NF-HEV, NFEHEV, C9orf26, NFHEV, Interleukin-33, Interleukin-1 family member 11, IL-33
Protein Accession Number:
NP_254274
Protein Gi Number:
15559209
Ncbi Gene Id Number:
90865
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
IL-33 Antibody: Interleukin-33 (IL-33) is a recently identified member of the IL-1 family of cytokines whose other members include IL-1α/beta, IL-1Ra and IL-18. Its receptor has been shown to be ST2, an IL-1 receptor family member that also acts as a negative regulator of TLR-IL-1R signaling and IL-1R accessory protein (IL-1RAcP). Receptor binding of IL-33 activates NF-κB and MAP kinases and induces the expression of TH2-associated cytokines such as IL-4, IL-5 and IL-6. Prolonged IL-33 treatment of mice led to the development of eosinophilia, splenomegaly, and severe pathological changes in mucosal organs such as lungs, esophagus and small intestine. Recent experiments have shown that IL-33 can also co-localize with heterochromatin and possesses transcriptional repressor activities, indicating that IL-33 may function as both a proinflammatory cytokine and an intracellular nuclear factor with transcriptional regulatory properties.
Background Reference 1:
Schmitz J et al. Immunity 2005; 23:479-90.
Background Reference 2:
Dinarello Eur. Cytokine Netw. 2000; 11:483-6.
Background Reference 3:
Brint et al. Nat. Immunol. 2004; 5:373-9.
Background Reference 4:
Chackerian et al. J. Immunol. 2007; 179:2551-5.
Citation 1:
Ivanov et al. Radiation-induced bystander signaling pathways in human fibroblasts: a role for interleukin-33 in the signal transmission.Cell Signal. 2010 Jul;22(7):1076-87. PMID: 20206688
Citation 2:
Tsuda et al. Novel splice variants of IL-33: differential expression in normal and transformed cells. J Invest Dermatol. 2012 Nov;132(11):2661-4. PMID: 22785132

FOR RESEARCH USE ONLY

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Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

4273

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

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