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5312
Human RANKL/TRANCE/TNFSF11 (hRANKL)
Cytokines & Growth Factors
Growth Factors and Cytokines

Human RANKL/TRANCE/TNFSF11 (hRANKL) #5312

Citations (0)
The induction of tartrate resistant acid phosphatase (TRAP) in Raw 264.7 cells was assessed. Raw 264.7 cells were treated with increasing concentrations of hRANKL for 4 days. Cells were lysed and TRAP activity of cell extracts was assessed and OD450 determined.
The purity of recombinant hRANKL was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hRANKL and staining overnight with Coomassie Blue.

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 hRANKL 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 hRANKL to surfaces. Solubilize for 30 minutes at room temperature with occasional gentle vortexing. Stock hRANKL 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) 30-35
Purity >98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hRANKL. All lots are greater than 98% pure.
Endotoxin Less than 0.01 ng endotoxin/1 μg hRANKL.
Activity The bioactivity of hRANKL was determined by measuring the ability of hRANKL to induce TRAP activity in Raw 264.7 cells. The ED50 of each lot is between 1.5-5 ng/ml.
Molecular Formula Recombinant hRANKL contains no "tags" and the nonglycosylated protein has a calculated MW of 20,484. DTT-reduced and non-reduced protein migrate as 30-35 kDa polypeptides. Lower mobility and heterogeneity in SDS-PAGE are due to glycosylation. The expected amino-terminal GSQHI of recombinant hRANKL was verified by amino acid sequencing.

Source / Purification

Recombinant human RANKL (hRANKL) Gly63-Asp244 (Accession #NP_143026) was expressed in human 293 cells at Cell Signaling Technology.

Background

RANKL, also known as TRANCE or OPGL, is a member of the TNF superfamily of ligands. T cells, mammary epithelial cells, and endothelial cells can produce RANKL (1). RANKL is expressed as a type II transmembrane protein or cleaved into a soluble form by extracellular proteases, such as TACE, ADAM10, and matrix metalloproteases (1). Alternative splicing also results in the production of soluble RANKL (1). RANKL signaling is antagonized by osteoprotegerin, which functions as a soluble decoy receptor (2). RANKL plays key roles in mammary gland development and dendritic cell survival and is required for osteoclast differentiation and survival (3-6). Research studies have shown that RANKL deficiencies in both mice and humans are associated with abnormally increased bone density and defects in lymphoid organogenesis (5,6).

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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To Purchase # 5312
Cat. # Size Qty. Price
5312SC
10 µg  (With Carrier)
5312SF
10 µg  (Carrier Free)