Product nameAnti-IL-1 beta antibody
See all IL-1 beta primary antibodies
DescriptionRabbit polyclonal to IL-1 beta
Tested applicationsSuitable for: WB, IP, ELISA, IHC-P, RIA, IHC-Fr, Functional Studies, Flow Cyt, Neutralising, ICC/IF, IHC-FoFrmore details
Species reactivityReacts with: Human
Predicted to work with: Non human primates
Recombinant 153 aa human IL-1beta produced in E.coli. MW of recombinant IL-1beta 17kDa, with the N-terminal amino acid at position alanine 117. This is the cleavage site generated by the IL-1beta converting enzyme (ICE, capase-1).
General notesImmunohistochemistry 1/100 to 1/200 (paraffin or cryofixation can be used; 1/100 for staining intracellular IL-1beta). ELISA 1/200 to 1/1000 (this antibody is best used as the second antibody with a monoclonal as a capture antibody). Radio immunoassay 1/8000 Neutralization of IL-1beta activity in bioassays 1/100 ( >4 hours incubation, normal rabbit IgG as negative control) FACS analysis - caution, F(c) domain of rabbit IgG may interact with cells non-specifically.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.20
Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
Concentration information loading...
Purification notesIgG from whole rabbit serum purified by DEAE fractionation.
Our Abpromise guarantee covers the use of ab2105 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration.
1/1000 - 1/2000 (antigen - supernatants or lysates of 2 x 106 endotoxin-stimulated human PBMC. Denatured 31kDa precursor IL-1beta will be recognized, but often migrates as a 35 kDa band).
|IP||Use at an assay dependent concentration. 1/400 - 1/800 (pre-clearing with a non-specific rabbit IgG is helpful to reduce background).|
|ELISA||1/500 - 1/2000.|
|IHC-P||1/100 - 1/200.|
|IHC-Fr||1/100 - 1/200.|
|Functional Studies||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.
F(c) domain of rabbit IgG may interact with cells non-specifically!
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
|Neutralising||1/100. Neutralization of IL-1beta activity in bioassays 1/100 ( >4 hours incubation, normal rabbit IgG as negative control). ab2105 does not neutralize the biological activity of murine, rat or rabbit IL1 beta.|
|ICC/IF||Use at an assay dependent concentration.|
|IHC-FoFr||Use at an assay dependent concentration. PubMed: 28726778|
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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ab2105 at 1/20 staining human liver tissue sections by IHC-P. The cells were paraformaldehyde fixed and a heat mediated antigen retrieval step was performed. The tissue was incubated with the tissue overnight at room temperature. A biotinylated donkey anti-rabbit IgG was used as the secondary antibody and this was detected using streptavidin HRP.
ab2105 staining IL-1 beta in human keratinocytes by Immunocytochemistry/ Immunofluorescence. Cells were fixed in methanol, permeabilized in 0,1% Saponin / PBS and then blocked using 4% BSA for 30 minutes at 25°C. Samples were then incubated with ab2105 at a 1/100 dilution for 2 hours at 4°C. An Alexa-fluor 488 conjugated goat anti-rabbit polyclonal was used as the secondary antibody at a 1/100 dilution.
All lanes : Anti-IL-1 beta antibody (ab2105) at 1/1000 dilution
Lane 1 : Human peripheral blood mononuclear cell lysate - PBMC's unstimulated
Lane 2 : Human peripheral blood mononuclear cell lysate - monocytes unstimulated
Lane 3 : Human peripheral blood mononuclear cell lysate - monocytes stimulated with LPS
Lane 4 : Human peripheral blood mononuclear cell lysate - positive control (rh IL-1 beta protein)
Lysates/proteins at 25 µg per lane.
All lanes : HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/2000 dilution
Performed under reducing conditions.
Observed band size: 35 kDa why is the actual band size different from the predicted?
Exposure time: 10 minutes
ab2105 staining IL-1 beta in Human monocytes by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with 0.1% saponin in PBS. The sample was incubated with the primary antibody (1/500 in 0.1% saponin + 1% FCS in PBS) for 45 minutes at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG (1/100) was used as the secondary antibody.
1 - unstimulated monocytes. 2 - monocytes stimulated with LPS.
Lanes 2-5 : Anti-IL-1 beta antibody (ab2105) at 1/1000 dilution
Lane 1 : Ladder (top to bottom: p35, p25, p20, p17, p11)
Lane 2 : Human THP-1 leukemic monocyte cell lysate, no LPS at 20 µg
Lane 3 : Human THP-1 leukemic monocyte supernatant, no LPS at 20 µg
Lane 4 : Human THP-1 leukemic monocyte cell lysate, LPS stimulation at 20 µg
Lane 5 : Human THP-1 leukemic monocyte supernatant, LPS stimulation at 20 µg
Lanes 2-5 : IRDye® 800CW-conjugated Donkey anti-rabbit IgG monoclonal at 1/5000 dilution
Performed under reducing conditions.
Observed band size: 17,30 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds
Both mature and pro-IL-1beta bands, p17 and p31 were detected by ab2105 in supernatant from THP-1 cells stimulated with LPS. Only pro-IL-1beta band (p31) was detected in supernatant from THP-1 cells not stimulated with LPS.
This product has been referenced in:
- Wang S et al. Oxidative stress, autophagy and pyroptosis in the neovascularization of oxygen-induced retinopathy in mice. Mol Med Rep 19:927-934 (2019). Read more (PubMed: 30569132) »
- Wang X et al. Effect of Gastrodin on Early Brain Injury and Neurological Outcome After Subarachnoid Hemorrhage in Rats. Neurosci Bull N/A:N/A (2019). Read more (PubMed: 30673960) »