IL-33 Antibody

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ProSci
Product Code: PSI-4273
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-4273-0.02mg0.02mg£150.00
Quantity:
PSI-4273-0.1mg0.1mg£449.00
Quantity:
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunofluorescence (IF)
  • Western Blot (WB)

Images

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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/mL Lane B: 2 μg/mL
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<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.
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<strong>Figure 3 Western Blot Validation of IL-33 in Human Lymph Node</strong><br> Loading: 15. μg of human lymph node lysate Antibodies: 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.
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<strong>Figure 4 Western Blot Validation of IL-33 in Human Lung</strong><br> Loading: 15. μg of human lung lysate Antibodies: 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.
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<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).
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<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.
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<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.
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<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.
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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/mL Lane 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 lysate Antibodies: 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 lysate Antibodies: 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.

Documents

Further Information

Additional Names:
IL-33 Antibody: DVS27, IL1F11, NF-HEV, NFEHEV, C9orf26, NFHEV, Interleukin-33, Interleukin-1 family member 11, IL-33
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.
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 References:
  • Schmitz J et al. Immunity 2005; 23:479-90.
  • Dinarello Eur. Cytokine Netw. 2000; 11:483-6.
  • Brint et al. Nat. Immunol. 2004; 5:373-9.
  • Chackerian et al. J. Immunol. 2007; 179:2551-5.
Buffer:
IL-33 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
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.
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.
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).
NCBI Gene ID #:
90865
NCBI Official Name:
interleukin 33
NCBI Official Symbol:
IL33
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 31kD

Observed: 33kD
Protein Accession #:
NP_254274
Protein GI Number:
15559209
Purification:
IL-33 Antibody is affinity chromatography purified via peptide column.
Research Area:
Immunology
Swissprot #:
O95760
User NOte:
Optimal dilutions for each application to be determined by the researcher.
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).

References

  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
  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

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