Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse

Applications

Validated:

Intracellular Staining by Flow Cytometry

Cited:

Flow Cytometry, Immunocytochemistry

Label

Phycoerythrin (Excitation = 488 nm, Emission = 565-605 nm)

Antibody Source

Monoclonal Rat IgG2A Clone # 396118
View all IL-33 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant mouse IL-33
Ser109-Ile266
Accession # Q8BVZ5

Specificity

Detects mouse IL-33 in direct ELISAs.

Clonality

Monoclonal

Host

Rat

Isotype

IgG2A

Scientific Data Images for Mouse IL‑33 PE‑conjugated Antibody

Detection of IL-33 antibody in bEnd.3 Mouse Cell Line antibody by Flow Cytometry.

Detection of IL‑33 in bEnd.3 Mouse Cell Line by Flow Cytometry.

bEnd.3 mouse endothelioma cell line was stained with Rat Anti-Mouse IL-33 PE-conjugated Mono-clonal Antibody (Catalog # IC3626P, filled histogram) or isotype control antibody (Catalog # IC006P, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Applications for Mouse IL‑33 PE‑conjugated Antibody

Application
Recommended Usage

Intracellular Staining by Flow Cytometry

10 µL/106 cells
Sample: bEnd.3 mouse endothelioma cell line fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

Reviewed Applications

Read 2 reviews rated 2 using IC3626P in the following applications:

Spectra Viewer

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

Shipping

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

Stability & Storage

Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

Background: IL-33

IL-33, also known as NF-HEV and DVS 27, is a 30 kDa proinflammatory protein that may also regulate gene transcription (1-3). DVS 27 was identifed as a gene that is upregulated in vasospastic cerebral arteries (1). NF-HEV was described as a nuclear factor that is preferentially expressed in the endothelial cells of high endothelial venules relative to endothelial cells from other tissues (2). IL-33 was identified based on sequence and structural homology with IL-1 family cytokines (3). DVS 27, NF-HEV, and IL-33 share 100% amino acid sequence identity. IL-33 is constitutively expressed in smooth muscle and airway epithelia. It is upregulated in arterial smooth muscle, dermal fibroblasts, and keratinocytes following IL-1 alpha or IL-1 beta stimulation (1, 3). Similar to IL-1, IL-33 can be cleaved in vitro by caspase-1, generating an N-terminal fragment that is slightly shorter than the C-terminal fragment (3, 4). The N-terminal portion of full length IL-33 contains a predicted bipartite nuclear localization sequence and a homeodomain-like helix-turn-helix DNA binding domain. By immunofluorescence, full length IL-33 localizes to the nucleus in HUVECs and transfectants (2). The C-terminal fragment, corresponding to mature IL-33, binds and triggers signaling through mast cell IL-1 R4/ST2L, a longtime orphan receptor involved in the augmentation of Th2 cell responses (3, 5-7). A ternary signaling complex is formed by the subsequent association of IL-33 and ST2L with IL-1R AcP (8). Stimulation of Th2 polarized lymphocytes with mature IL-33 in vitro induces IL-5 and IL-13 secretion (3). In vivo administration of mature IL-33 promotes increased production of IL-5, IL-13, IgE, and IgA, as well as splenomegaly and inflammatory infiltration of mucosal tissues (3). Full length and mature mouse IL-33 share approximately 55% and 90% amino acid (aa) sequence identity with human and rat IL-33, respectively. Mouse IL-33 shares less than 25% aa sequence identity with other IL-1 family proteins.

References

  1. Onda, H. et al. (1999) J. Cereb. Blood Flow Metab. 19:1279.
  2. Baekkevold, E.S. et al. (2003) Am. J. Pathol. 163:69.
  3. Schmitz, J. et al. (2005) Immunity 23:479.
  4. Black, R.A. et al. (1989) J. Biol. Chem. 264:5323.
  5. Xu, D. et al. (1998) J. Exp. Med. 187:787.
  6. Lohning, M. et al. (1998) Proc. Natl. Acad. Sci. USA 95:6930.
  7. Dinarello, C.A. (2005) Immunity 23:461.
  8. Chackerian, A.A. et al. (2007) J. Immunol. 179:2551.
    9.  

Long Name

Interleukin 33

Alternate Names

C9orf26, DVS27, IL33, NF-HEV

Entrez Gene IDs

90865 (Human); 77125 (Mouse)

Gene Symbol

IL33

UniProt

Q8BVZ5

Additional IL-33 Products

Product Documents for Mouse IL‑33 PE‑conjugated Antibody

Certificate of Analysis

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Product Specific Notices for Mouse IL‑33 PE‑conjugated Antibody

For research use only

Citations for Mouse IL‑33 PE‑conjugated Antibody

Customer Reviews for Mouse IL‑33 PE‑conjugated Antibody (2)

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  • Mouse IL-33 PE-conjugated Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Lung single-cell suspension
    Species: Mouse
    Verified Customer | Posted 08/30/2016
    Cells were activated with PMA and ionomycin for 2 hrs before intracellular staining.
    Mouse IL‑33 PE‑conjugated Antibody IC3626P
  • Mouse IL-33 PE-conjugated Antibody
    Name: Oliver Burton
    Application: Flow Cytometry
    Sample Tested: mast cells
    Species: Mouse
    Verified Customer | Posted 07/25/2016
    Mast cells were cultured from the bone marrow of IL-33-deficient or WT C57BL/6 mice. Genotypes were confirmed by PCR. Cells (5x10^5) were stained for viability, FceRI and c-kit, blocking Fc receptors, then fixed with BD Cytofix/Cytoperm. Cells were permeabilized with BD Permeabilization Buffer, and stained with 2ul anti-mouse IL-33-PE or rat IgG2a-PE isotype control. Cells were washed three times, and data were acquired on a BD FACSCanto LSR flow cytometer, gating on live c-Kit+FceRI+ cells. Fluorescence increases by two orders of magnitude in both WT and IL-33-KO cells following staining with the IL-33-PE antibody relative to the isotype control. I am currently investigating whether there is any specific signal after stimulation of the cells, at which point IL-33 is induced. Staining with the recommended (greater) amount of antibody produces an even greater background shift in unstimulated cells.
    Mouse IL‑33 PE‑conjugated Antibody IC3626P
    Bio-Techne Response
    R&D Systems Technical Service is investigating.

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