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The production of mouse IL-6 by 3T3 MEFs WT cultured with increasing concentrations of mIL-1β was assessed. Media from cells incubated with mIL-1β for 24 hours was collected and assayed for mouse IL-6 by ELISA and the OD450-OD650 was determined.
The purity of recombinant mIL-1β was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mIL-1β and staining overnight with Coomassie Blue.
Western blot analysis of extracts from 3T3 MEFs WT untreated or treated with mIL-1β for 15 minutes, using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033 (upper) and NF-κB p65 (E498) Antibody #3987 (lower).
Product Information
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg mIL-1β. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.
Stable in lyophilized state at 4°C for 1 year after receipt. Sterile stock solutions reconstituted with carrier protein are stable at 4°C for 2 months and at -20°C for 6 months. Avoid repeated freeze-thaw cycles.Maintain sterility. Storage at -20°C should be in a manual defrost freezer.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mIL-1β. All lots are greater than 98% pure.
Recombinant mIL-1β has a Met on the amino terminus and has a calculated MW of 17,525. DTT-reduced and non-reduced protein migrate as 18 kDa polypeptides. The expected amino-terminal MVPIR of recombinant mIL-1β was verified by amino acid sequencing.
The bioactivity of recombinant mIL-1β was determined by its ability to induce mouse IL-6 production by 3T3 MEFs WT. The ED50 of each lot is between 5-20 pg/ml.
Less than 0.01 ng endotoxin/1 μg mIL-1β.
Recombinant mouse IL-1β (mIL-1β) Val118-Ser269 (Accession #NP_032387) was produced in E.coli at Cell Signaling Technology.
IL-1β is a pro-inflammatory cytokine produced predominantly by activated monocytes and epithelial cells (1). Precursor IL-1β is cleaved by caspase-1 and mature IL-1β is then secreted (1-3). Target cells include macrophages and many other cell types. Signaling by IL-1β involves IL-1β binding to IL-1 accessory protein (IL-1-AcP) and then the complex binds to IL-1RI (1,2). Signaling is through activation of MAP kinase and NF-κB pathways (1,2). IL-1β also binds to IL-1RII that lacks an intracellular signaling domain and thereby serves as a high affinity decoy receptor. IL-1β binding to IL-1RI is inhibited by the negative regulator, IL-1R antagonist (IL-1Ra). IL-1Ra binding to IL-1RI does not signal and serves to block IL-1β signaling. IL-1β plays critical roles in the acute phase response and sepsis (1-3).
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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
| Product # | Size | Price |
|---|---|---|
| 5204SC | 10 µg (With Carrier) | $ 221 |
| 5204SF | 10 µg (Carrier Free) | $ 221 |
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