Product nameAnti-IL-1 beta antibody [EPR16805-15]
See all IL-1 beta primary antibodies
DescriptionRabbit monoclonal [EPR16805-15] to IL-1 beta
Tested applicationsSuitable for: WB, Flow Cyt, IPmore details
Unsuitable for: ICC/IF or IHC-P
Species reactivityReacts with: Mouse
Recombinant fragment within Mouse IL-1 beta aa 100 to the C-terminus. The exact sequence is proprietary.
Database link: P10749
- WB: RAW 264.7 treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, then with 300 ng/ml Brefeldin A (BFA) added after 3 hours, whole cell lysate. IP: RAW 264.7 treated with 100 ng/ml LPS for 3h, then add 300 ng/ml Brefeldin A for 3h, whole cell lysate. Flow cyt: RAW 264.7 cells treated with 100 ng/ml LPS for 3h, then add 300 ng/ml Brefeldin A for 3h.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab234437 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Detects a band of approximately 31, 28, 17 kDa (predicted molecular weight: 30 kDa).|
FunctionPotent proinflammatory cytokine. Initially discovered as the major endogenous pyrogen, induces prostaglandin synthesis, neutrophil influx and activation, T-cell activation and cytokine production, B-cell activation and antibody production, and fibroblast proliferation and collagen production. Promotes Th17 differentiation of T-cells.
Tissue specificityExpressed in activated monocytes/macrophages (at protein level).
Sequence similaritiesBelongs to the IL-1 family.
modificationsActivation of the IL1B precursor involves a CASP1-catalyzed proteolytic cleavage. Processing and secretion are temporarily associated.
Cellular localizationCytoplasm, cytosol. Lysosome. Secreted, exosome. Cytoplasmic vesicle, autophagosome. Secreted. The precursor is cytosolic. In response to inflammasome-activating signals, such as ATP for NLRP3 inflammasome or bacterial flagellin for NLRC4 inflammasome, cleaved and secreted. IL1B lacks any known signal sequence and the pathway(s) of its secretion is(are) not yet fully understood (PubMed:24201029). On the basis of experimental results, several unconventional secretion mechanisms have been proposed. 1. Secretion via secretory lysosomes: a fraction of CASP1 and IL1B precursor may be incorporated, by a yet undefined mechanism, into secretory lysosomes that undergo Ca(2+)-dependent exocytosis with release of mature IL1B (PubMed:15192144). 2. Secretory autophagy: IL1B-containing autophagosomes may fuse with endosomes or multivesicular bodies (MVBs) and then merge with the plasma membrane releasing soluble IL1B or IL1B-containing exosomes (PubMed:24201029). However, autophagy impacts IL1B production at several levels and its role in secretion is still controversial. 3. Secretion via exosomes: ATP-activation of P2RX7 leads to the formation of MVBs containing exosomes with entrapped IL1B, CASP1 and other inflammasome components. These MVBs undergo exocytosis with the release of exosomes. The release of soluble IL1B occurs after the lysis of exosome membranes (By similarity). 4. Secretion by microvesicle shedding: activation of the ATP receptor P2RX7 may induce an immediate shedding of membrane-derived microvesicles containing IL1B and possibly inflammasome components. The cytokine is then released in the extracellular compartment after microvesicle lysis (PubMed:11728343). 5. Release by translocation through permeabilized plasma membrane. This may occur in cells undergoing pyroptosis due to sustained activation of the inflammasome (By similarity). These mechanisms may not be not mutually exclusive.
- Information by UniProt
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IL-1 beta was immunoprecipitated from 0.35 mg of RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) (treated with 100 ng/ml LPS for 3h, then add 300 ng/ml Brefeldin A for 3h) whole cell lysate with ab234437 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab234437 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.
Lane 1: RAW 264.7 (treated with 100 ng/ml LPS for 3h, then add 300 ng/ml Brefeldin A for 3h) whole cell lysate 10 μg (Input).
Lane 2: ab234437 IP in RAW 264.7 (treated with 100 ng/ml LPS for 3h, then add 300 ng/ml Brefeldin A for 3h) whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab234437 in RAW 264.7 (treated with 100 ng/ml LPS for 3h, then add 300 ng/ml Brefeldin A for 3h) whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
Flow cytometric 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 that were either treated (100 ng/ml LPS for 3h, then add 300 ng/ml Brefeldin A for 3h labeling)(red) or untreated (green) labeling IL-1 beta with ab234437 at 1/60 dilution compared with a rabbit monoclonal IgG Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti-Rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
All lanes : Anti-IL-1 beta antibody [EPR16805-15] (ab234437) at 1/1000 dilution
Lane 1 : Untreated RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 2 : RAW 264.7 (treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, then with 300 ng/ml Brefeldin A (BFA) added after 3 hours) whole cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 30 kDa
Observed band size: 17,28,31 kDa why is the actual band size different from the predicted?
Exposure time: 15 seconds
Blocking and dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 8446594).
ab234437 has not yet been referenced specifically in any publications.