The viability of BAF3 cells treated with increasing amounts of mLCN2 was assessed. After 48 hr treatment with mLCN2, cells were incubated with a tetrazolium salt and the OD450-OD650 was determined.
The purity of recombinant mLCN2 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mLCN2 and staining overnight with Coomassie Blue.
With carrier: A 0.22 μm filtered solution of 0.33 mg/ml mLCN2 in 20 mM Tris, pH 8.0 containing 150 mM NaCl and 20 μg BSA per 1 μg mLCN2. Carrier free: A 0.22 μm filtered solution of 0.33 mg/ml mLCN2 in 20 mM Tris, pH 8.0 containing 150 mM NaCl.
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 mLCN2 All lots are greater than 98% pure.
Recombinant mLCN2 contains no "tags" and the nonglycosylated protein has a calculated MW of 20,874. DTT-reduced and non-reduced proteins migrate as 24 kDa polypeptides.
The bioactivity of mLCN2 was determined in a BAF3 cell viability assay. The ED50 of each lot is between 0.5-4 µg/ml.
Less than 0.01 ng endotoxin/1 μg mLCN2.
Recombinant mouse LCN2 (mLCN2) Gln21-Asn200 (Accession #P11672) was expressed in human 293 cells at Cell Signaling Technology.
Lipocalin-2, a member of the lipocalin family of proteins, was originally identified as a gelatinase-associated component of neutrophil secretory granules (1). Lipocalin-2 is involved in innate immunity, iron homeostasis, and apoptosis. Lipocalin-2 limits bacterial growth by binding to bacterial siderophores and sequestering iron (2-4). In mammalian cells, iron-loaded lipocalin-2 binds to its receptor, 24p3R, and is internalized, thereby releasing iron and increasing the intracellular iron concentration (5). Conversely, iron-free lipocalin-2 promotes apoptosis (5). Lipocalin-2 is also expressed in adipose tissue and promotes insulin resistance in cultured mouse adipocytes (6).
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