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5216
Mouse Interleukin-6 (mIL-6)
Cytokines & Growth Factors
Growth Factors and Cytokines

Mouse Interleukin-6 (mIL-6) #5216

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# Product Name Applications Reactivity
The proliferation of B9 cells treated with increasing concentrations of mIL-6 was assessed. After 48 hour treatment with mIL-6, cells were incubated with a tetrazolium salt and the OD450 - OD650 was determined.
The purity of recombinant mIL-6 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mIL-6 and staining overnight with Coomassie Blue.
Western blot analysis of extracts from B9 cells, untreated or treated with increasing concentrations of mIL-6 for 10 minutes, using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145 (upper) and Stat3 (79D7) Rabbit mAb #4904 (lower).

Product Usage Information

Reconstitution:

With carrier: Add sterile PBS or PBS containing 1% bovine or human serum albumin or 5-10% FBS to a final mIL-6 concentration of greater than 50 μg/ml. Solubilize for 30 minutes at room temperature with occasional gentle vortexing.

Carrier free: Add sterile PBS or PBS containing protein to minimize absorption of mIL-6 to surfaces. Solubilize for 30 minutes at room temperature with occasional gentle vortexing. Stock mIL-6 should be greater than 50 μg/ml.

Storage

Stable in lyophilized state at -20°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.

Product Description

MW (kDa) 26-36
Purity >98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mIL-6. All lots are greater than 98% pure.
Endotoxin Less than 0.01 ng endotoxin/1 μg mIL-6.
Activity The bioactivity of recombinant mIL-6 was determined in a B9 cell proliferation assay. The ED50 of each lot is between 0.5 and 20 pg/ml.
Molecular Formula Recombinant mIL-6 contains no "tags" and the nonglycosylated protein has a calculated MW of 21,734. DTT-reduced and non-reduced protein migrate as 31 kDa polypeptides. Lower mobility in SDS-PAGE is due to glycosylation. The expected amino-terminal FPTSQ of recombinant mIL-6 was verified by amino acid sequencing.

Source / Purification

Recombinant mouse IL-6 (mIL-6) Phe25-Thr211 (Accession #NP_112445) was expressed in human 293 cells at Cell Signaling Technology.

Background

IL-6 is a potent inducer of the acute phase response and is produced by T cells, macrophages, fibroblasts, endothelial, and other cells (1,2). IL-6 induces proliferation and differentiation and acts on B cells, T cells, thymocytes, and others. IL-6 in concert with TGFβ is important for developing Th17 responses. IL-6 binds to IL-6Rα and through this association induces gp130 homodimerization (1). gp130 homodimerization triggers the Jak/Stat cascade and the SHP2/MAPK (Erk) cascade (1,3,4). IL-6 also forms a complex with an IL-6Rα splice variant that is non-membrane associated (3).The IL-6/soluble IL-6Rα complex can then activate the gp130 signaling pathway on cells that express gp130 but not IL-6Rα (3). IL-6, through increasing expression of proangiogenic VEGF, may contribute to metastatic breast cancer (5).

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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