Recombinant Rat RANKL (TRANCE, TNFSF11) (carrier-free)

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Other Names
Tumor necrosis factor ligand superfamily, member 11 (TNFSF11), Osteoprotegerin ligand (OPGL), Receptor activator of NF-kappa-B ligand (RANKL), TNF-related activation-induced cytokine (TRANCE), Osteoclast differentiation factor (ODF), CD254
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Product Citations
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Rat_RANKL_CF_RECOM_1_031519
Rat RANKL induces osteoclast differentiation in mouse macrophage RAW264.7 cell line. The ED50 for this effect is 2 – 10 ng/mL.
  • Rat_RANKL_CF_RECOM_1_031519
    Rat RANKL induces osteoclast differentiation in mouse macrophage RAW264.7 cell line. The ED50 for this effect is 2 – 10 ng/mL.
  • Rat_RANKL_CF_RECOM_2_031519
    Stability Testing for Recombinant Rat RANKL. Recombinant Rat RANKL was aliquoted in in 20 mM sodium phosphate, pH 6.0, 100 mM NaCl, and 2 mM EDTA at 0.2 mg/mL. One aliquot was frozen and thawed four times (4x Freeze/Thaw) and compared to the control that was kept at 4°C (control). The samples were tested for their ability to induce osteoclast differentiation in RAW264.7 cells. The ED50 for this effect is 2 – 10 ng/mL.
Cat # Size Price Quantity Avail. Save
779202 10 µg 125 CHF
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779204 25 µg 245 CHF
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779206 100 µg 730 CHF
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Description

The RANKL gene encodes a type II membrane protein of 316 amino acids with a predicted molecular mass of 35 kD. RANKL is cleaved to produce a soluble form with biological activity. The shedding of membrane-bound RANKL appears to be mediated by expression of MMP14 and ADAM10. Suppression of MMP14 in primary osteoblasts increases membrane-bound RANKL and promotes osteoclastogenesis in cocultures with macrophages. Therefore, RANKL shedding seems to be an important process that downregulates local osteoclastogenesis. Alternatively, an increased production of RANKL by osteoblastic cells leads to osteoclast differentiation, activation, and survival, which results in increased bone resorption. Binding of RANKL to its receptor RANK activates TNF receptor-associated factor 6 (TRAF6), which is linked to downstream pathways including NF-κB, c-jun N-terminal kinase (JNK) or Src. TRAF6 has been shown to be necessary for the differentiation of osteoclastic cells by enhancing Src kinase, essential for osteoclast function.

Product Details
Technical Data Sheet (pdf)

Product Details

Source
Rat RANKL, amino acid (Arg72-Asp318) (Accession: #Q9ESE2.1), with an N-terminal 6His tag, was expressed in CHO cells.
Molecular Mass
The 253 amino acid recombinant protein has a predicted molecular mass of approximately 28.65 kD. The DTT-reduced and non-reduced protein migrates at approximately 33 kD respectively by SDS-PAGE. The predicted N-terminal amino acid is His.
Purity
>95%, as determined by Coomassie stained SDS-PAGE.
Formulation
0.22 µm filtered protein solution is in 20 mM sodium phosphate, pH 6.0, 100 mM NaCl and 2 mM EDTA.
Endotoxin Level
Less than 1.0 EU per µg protein as determined by the LAL method.
Concentration
10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg and larger sizes are lot-specific and bottled at the concentration indicated on the vial (please contact technical support for concentration, or use our Lookup tool if you have a lot number.)
Storage & Handling
Unopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
Activity
The ED50= 2 - 10 ng/mL, as determined by induction of osteoclast differentiation in mouse macrophage RAW264.7 cell line, measured by TRAP reaction.
Application

Bioassay

Application Notes

BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com.

Antigen Details

Structure
Cytokine
Distribution

Predominantly expressed in monocytes, macrophages, polymorphonuclear cells, B cells. Detected in microglial cells and Treg cells. Inducible in keratinocytes and endothelial cells.

Function
Potent stimulator of osteoclast formation from precursor cells and bone-resorbing activity in mature osteoclasts. Augments the ability of dendritic cells to stimulate naïve T-cell proliferation. Activates JNK and NF-κB and induces the expression of IL-1, IL-6, IL-12, and IL-15 in dendritic cells. Crucial factor in lymphocyte development and lymph node organogenesis. Stimulates proliferation, adhesion, and IL-7 expression of thymic epithelial cells. RANKL expression, in human and murine osteoblastic cells, is stimulated by various cytokines (IL-1, TNF-α, and IL-11) and calciotrophic hormones including PTH, 1, 25-dihydroxyvitamin D3 (1,25D3), and prostaglandin E2.
Interaction
T cells, dendritic cells, bone marrow derived macrophages, osteoclast precursors and mature osteoclasts.
Ligand/Receptor
TNFRSF11A (Tumor necrosis factor receptor superfamily, member 11A) also called RANK and TNFRSF11B (OPG) which acts as a decoy receptor for RANKL and as a potent inhibitor of osteoclast formation.
Bioactivity
Rat RANKL induces osteoclast differentiation in RAW264.7 cells.
Cell Type
Dendritic cells, Osteoclasts, T cells, Embryonic Stem Cells
Biology Area
Cell Biology, Immunology, Signal Transduction, Stem Cells
Molecular Family
Cytokines/Chemokines, Growth Factors
Antigen References
  1. Dehm SM, et al. 2004. Biochem. Cell Biol. 82:263.
  2. Hikita A, et al. 2006. J. Biol. Chem. 281:36846.
  3. Kim NS, et al. 2006. Mol. Cell. Biol. 26:1002.
  4. Wada T, et al. 2006. TRENDS Mol. Med. 12:17.
  5. Lee HW, et al. 2008. Exp. Mol. Med. 40:59.
  6. Ha J, et al. 2010. J. Immunol. 184:4717.
Gene ID
117516 View all products for this Gene ID
UniProt
View information about RANKL on UniProt.org

Related FAQs

Does specific activity of a recombinant protein vary between lots?

Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.

Have your recombinants been tested for stability?

Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.

How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
Use formula Specific activity (Units/mg) = 10e6/ ED50 (ng/mL)
How does the activity of your recombinant proteins compare to competitors?

We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!

What is the specific activity or ED50 of my recombinant protein?

The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.

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For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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