Product Pathways - Jak/Stat Pathway
Cleaved IL-1β (Asp116) Antibody #2021
PhosphoSitePlus® protein, site, and accession data: IL1B
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W | H | Endogenous | 17 | Rabbit |
Applications Key:
W=Western Blotting
Reactivity Key:
H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 2021:
- Western Blotting
Specificity / Sensitivity
Cleaved IL-1beta (Asp116) Antibody detects endogenous levels of the 17 kDa processed form of IL-1beta. The antibody does not recognize the uncleaved precursor of IL-1beta.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino-terminal sequence of the 17 kDa mature form of human IL-1beta. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts (lanes 1 and 2) and supernatants (lanes 3 and 4) from THP-1 cells, untreated or lipopolysaccharide (LPS)-treated (10 ug/ml for 24 hours), using Cleaved IL-1beta (Asp116) Antibody (upper) or IL-1beta Antibody (lower). Cells were differentiated with 0.5 uM TPA for 3 hours. Cells were washed and plated onto 24-well plates at a density of 4 x 105 cells per well and left to adhere overnight prior to LPS treatment with the addition of 5 mM ATP for 1 hour to stimulate release of IL-1 beta.
Background
Interleukin-1beta (IL-1beta), one of the major caspase-1 targets, is a multifunctional cytokine that is involved in a host of immune and proinflammatory responses (1). It is produced primarily by activated monocytes and macrophages. It signals through various adaptor proteins and kinases that lead to activation of numerous downstream targets (2-6). Human IL-1beta is synthesized as a 31 kDa precursor. To gain activity, the precursor must be cleaved by caspase-1 between Asp116 and Ala117 to yield a 17 kDa mature form (7,8). Detection of the 17 kDa mature form of IL-1beta is a good indicator of caspase-1 activity.
- Dinarello, C.A. (1998) Int. Rev. Immunol. 16, 457-499.
- Burns, K. et al. (1998) J. Biol. Chem. 273, 12203-12209.
- Cao, Z. et al. (1996) Nature 383, 443-446.
- Cao, Z. et al. (1996) Science 271, 1128-1131.
- Wesche, H. et al. (1997) Immunity 7, 837-847.
- Ninomiya-Tsuji, J. et al. (1999) Nature 398, 252-256.
- Thornberry, N. A. et al. (1992) Nature 356, 768-774.
- Cerretti, D. P. et al. (1992) Science 256, 97-100.
- Grahames, C.B. et al. (1999) Br J Pharmacol 127, 1915-21.
Application References
- Martinon, F. et al. (2002) The Inflammasome: A Molecular Platform Triggering Activation Of Inflammatory Caspases and processing of proIL-beta. Molecular Cell 10, 417-426. Applications: Western Blotting
- Lee, J. et al. (2011) Oncogene , . Applications: Western Blotting
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- 7003 20X LumiGLO® Reagent and 20X Peroxide
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
