The proliferation of 2E8 cells treated with increasing concentrations of mIL-7 was assessed. After 48 hour treatment with
mIL-7, cells were incubated with a tetrazolium salt and the OD450 - OD650 was determined.
The purity of recombinant mIL-7 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mIL-7 and staining overnight with Coomassie Blue.
Western blot analysis of extracts from 2E8 cells, untreated or treated with mIL-7 for 15 minutes, using Phospho-Stat5 (Tyr695) (C11C5) Rabbit mAb Antibody #9359 (upper) and Stat5 (3H7) Antibody #9358 (lower).
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg mIL-7. 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-7. All lots are greater than 98% pure.
Recombinant mIL-7 has a Met on the amino terminus and has a calculated MW of 15,028. DTT-reduced and non-reduced protein migrate as 14 kDa polypeptides. The expected amino-terminal MECHI of recombinant mIL-7 was verified by amino acid sequencing.
The bioactivity of recombinant mIL-7 was determined in a 2E8 cell proliferation assay. The ED50 of each lot is between 50-500 pg/ml.
Less than 0.01 ng endotoxin/1 μg mIL-7.
Recombinant mouse IL-7 (mIL-7) Glu26-Ile154 (Accession #NP_032397) was produced in E. coli at Cell Signaling Technology.
IL-7 plays a key role in lymphopoiesis and lymphoid homeostasis (1). Stromal and epithelial cells within the bone marrow and thymus produce IL-7 (1). The primary targets of IL-7 are T cells, B cells, and dendritic cells (1). IL-7 is crucial for T cell development, the importance of which is underscored by the lack of T cells in both mice and humans that are deficient in IL-7/IL-7R signaling (1,2). While IL-7 appears to be required for B cell development in mice, the role of IL-7 in human B cell development is unclear (1,3). In addition to its effects on T cell lymphopoiesis, IL-7 promotes the maintenance and survival of naïve and memory αβ T cells, as well as γδ T cells (1). The IL-7 receptor is a heterodimer of the common γ chain, γc, and the IL-7-specific IL-7Rα (1). IL-7 activates PI3K/Akt, Jak1/2, and Stat1, 3, and 5 (1).
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