Recombinant Anti-IL-33 antibody [EPR17831] - BSA and Azide free (ab229698)
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 - BSA and Azide free
- 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] - BSA and Azide free
See all IL-33 primary antibodies -
Description
Rabbit monoclonal [EPR17831] to IL-33 - BSA and Azide free -
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
- IHC: Mouse spleen tissue. Flow Cyt (intra): RAW 264.7 cells
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General notes
ab229698 is the carrier-free version of ab187060.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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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Conjugation kits
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab229698 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 |
Use at an assay dependent concentration. Detects a band of approximately 33 kDa (predicted molecular weight: 30 kDa).
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IHC-P |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
Use at an assay dependent concentration. Detects a band of approximately 33 kDa (predicted molecular weight: 30 kDa). |
IHC-P
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
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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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187060).
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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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187060).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187060).
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Intracellular Flow Cytometry analysis ofRAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 50nM PMA and 5�g/ml LPS for 24h (Red) / Untreated control (Green) labeling IL-33 with ab229698 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187060).
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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Datasheet download
Certificate of Compliance
References (2)
ab229698 has been referenced in 2 publications.
- Chen L et al. Tumor-Derived IL33 Promotes Tissue-Resident CD8+ T Cells and Is Required for Checkpoint Blockade Tumor Immunotherapy. Cancer Immunol Res 8:1381-1392 (2020). PubMed: 32917659
- He Z et al. Mast cells are essential intermediaries in regulating IL-33/ST2 signaling for an immune network favorable to mucosal healing in experimentally inflamed colons. Cell Death Dis 9:1173 (2018). PubMed: 30518915