The purity of recombinant mOSM was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mOSM and staining overnight with Coomassie Blue.
The proliferation of NIH/3T3 cells treated with increasing concentrations of mOSM was assessed. After 24 hr treatment, cells were labeled with BrdU for 4 hrs. BrdU incorporation was determined by ELISA and the OD450-OD690 was determined.
Western blot analysis of extracts from MEF cells untreated or treated with mOSM for 15 minutes, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (upper) and p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg mOSM. 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 mOSM. All lots are greater than 98% pure.
Recombinant mOSM does not have Met on the amino terminus and has a calculated MW of 20,750. DTT-reduced and non-reduced protein migrate as 21 kDa polypeptides. The expected amino-terminal ANRGC of recombinant mOSM was verified by amino acid sequencing.
The bioactivity of recombinant mOSM was determined in a NIH/3T3 cell proliferation assay. The ED50 of each lot is between 0.10-0.40 ng/ml.
Less than 0.01 ng endotoxin/1 μg mOSM.
Recombinant mouse OSM (mOSM) Ala24-Arg206 (Accession #NP_001013383) was produced in E. coli at Cell Signaling Technology.
Oncostatin M (OSM) is a member of the IL-6 family of cytokines that is produced primarily by activated T cells and macrophages (1,2). OSM induces fibroblast proliferation, inhibits tumor cell proliferation, and plays a role in immune regulation (2,3). Mouse OSM binds to the OSM receptor β (OSMRβ) which forms a heteromeric complex with the common IL-6 family receptor subunit, gp130 (4). In contrast, human OSM binds to two distinct receptor complexes, the OSMRβ/gp130 and LIFRβ/gp130 complexes (4). The ability of human OSM to bind to two receptors may explain the overlapping and distinct activities of human OSM and LIF. OSM induces the activation the JAK2, STAT3, and ERK1/2 pathways (5).
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