Recombinant Anti-IL-33 antibody [EPR17831] (ab187060)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17831] to IL-33
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-IL-33 antibody [EPR17831]
See all IL-33 primary antibodies -
Description
Rabbit monoclonal [EPR17831] to IL-33 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Rat and mouse lung tissue lysate; RAW 264.7 treated with 50 nM Phorbol-12-myristate-13-acetate (PMA) and 5 ug/ml lipopolysaccharide (LPS) for 24 hours, whole cell lysate. IHC-P: Mouse and rat spleen tissue. ICC/IF: RAW 264.7 treated with 50 nM Phorbol-12-myristate-13-acetate (PMA) and 5 ug/ml lipopolysaccharide (LPS) for 24 hours cells. Flow Cyt (intra): RAW 264.7 cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17831 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab187060 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/500.
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WB |
1/1000. Detects a band of approximately 33 kDa (predicted molecular weight: 30 kDa).
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IHC-P | (1) |
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Antigen retrieval performed using Universal HIER antigen retrieval reagent (10X) (ab208572). |
ICC/IF | (1) |
1/500.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Detects a band of approximately 33 kDa (predicted molecular weight: 30 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Antigen retrieval performed using Universal HIER antigen retrieval reagent (10X) (ab208572). |
ICC/IF
1/500. |
Target
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Function
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 (PubMed:16286016). 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 (PubMed:17853410, PubMed:18836528).
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 (PubMed:21734074). This form is rapidely lost upon angiogenic or proinflammatory activation (PubMed:18787100). -
Tissue specificity
Expressed at high level in high endothelial venules found in tonsils, Peyer patches and mesenteric lymph nodes. Almost undetectable in placenta. -
Sequence similarities
Belongs to the IL-1 family. Highly divergent. -
Domain
The homeodomain-like HTH domain mediates nuclear localization and heterochromatin association. -
Post-translational
modificationsThe full length protein can be released from cells and is able to signal via the IL1RL1/ST2 receptor. However, proteolytic processing by CSTG/cathepsin G and ELANE/neutrophil elastase produces C-terminal peptides that are more active than the unprocessed full length protein. May also be proteolytically processed by calpains (PubMed:19596270). Proteolytic cleavage mediated by apoptotic caspases including CASP3 and CASP7 results in IL33 inactivation (PubMed:19559631). In vitro proteolytic cleavage by CASP1 was reported (PubMed:16286016) but could not be confirmed in vivo (PubMed:19465481) suggesting that IL33 is probably not a direct substrate for that caspase. -
Cellular localization
Nucleus. Chromosome. Cytoplasmic vesicle, secretory vesicle. Secreted. Associates with heterochromatin and mitotic chromosomes (PubMed:17185418). - Information by UniProt
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Database links
- Entrez Gene: 77125 Mouse
- Entrez Gene: 361749 Rat
- SwissProt: Q8BVZ5 Mouse
- SwissProt: Q66H70 Rat
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Alternative names
- C9orf26 antibody
- CHROMOSOME 9 OPEN READING FRAME 26 antibody
- DKFZp586H0523 antibody
see all
Images
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Flow cytometry overlay histogram showing left, Raw264.7 treated with 50nM PMA and 5μg/ml LPS for 24h and right, negative untreated Raw264.7 stained with ab187060 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab187060) (1x 106 in 100μl at 1.0μg/ml (1/2090)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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All lanes : Anti-IL-33 antibody [EPR17831] (ab187060) at 1/1000 dilution
Lane 1 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 2 : RAW 264.7 treated with 50 nM Phorbol-12-myristate-13-acetate (PMA) and 5 ug/ml lipopolysaccharide (LPS) for 24 hours, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Developed using the ECL technique.
Predicted band size: 30 kDa
Observed band size: 33 kDa why is the actual band size different from the predicted?Blocking/Dilution: 5% NFDM/TBST
Exposure: 3 minutes
IL-33 expression is induced by LPS treatment of PMA-differentiated RAW 264.7 cells (PMID 19559631; PMID 19933859).
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All lanes : Anti-IL-33 antibody [EPR17831] (ab187060) at 1/1000 dilution
Lane 1 : Rat lung tissue lysate at 20 µg
Lane 2 : Mouse lung tissue lysate at 10 µg
Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 30 kDa
Observed band size: 30 kDaBlocking/Dilution: 5% NFDM/TBST
Exposure: 3 minutes
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Intracellular Flow Cytometry analysis ofRAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 50nM PMA and 5µg/ml LPS for 24h (Red) / Untreated control (Green) labeling IL-33 with ab187060 at 1/500 dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody. Cells were fixed with4% paraformaldehyde and permeabilised with0.1% Tween-20.Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling IL33 with ab187060 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use. Nuclear staining in endothelial cells of mouse spleen is observed (PMID: 12819012). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling IL33 with ab187060 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use. Nuclear staining in endothelial cells of rat spleen is observed (PMID: 12819012). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling IL33 with ab187060 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing increased nuclear staining in RAW 264.7 cells treated with 50 nM Phorbol-12-myristate-13-acetate (PMA) and 5 µg/ml Lipopolysaccharide for 24h.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (11)
ab187060 has been referenced in 11 publications.
- Van Mechelen M et al. Impact of barrier tissue inflammation and physical activity on joint homeostasis in mice. Rheumatology (Oxford) 61:1690-1698 (2022). PubMed: 34175921
- Wang Z et al. KIF2A decreases IL-33 production and attenuates allergic asthmatic inflammation. Allergy Asthma Clin Immunol 18:55 (2022). PubMed: 35718777
- He X et al. Akkermansia muciniphila Alters Gut Microbiota and Immune System to Improve Cardiovascular Diseases in Murine Model. Front Microbiol 13:906920 (2022). PubMed: 35774450
- Cheng L et al. IL-33 Deficiency Attenuates Lung Inflammation by Inducing Th17 Response and Impacting the Th17/Treg Balance in LPS-Induced ARDS Mice via Dendritic Cells. J Immunol Res 2022:9543083 (2022). PubMed: 36570798
- Deng F et al. Gut Microbial Metabolite Pravastatin Attenuates Intestinal Ischemia/Reperfusion Injury Through Promoting IL-13 Release From Type II Innate Lymphoid Cells via IL-33/ST2 Signaling. Front Immunol 12:704836 (2021). PubMed: 34650552