RANKL/TNFSF11 Protein, Human (HEK293)

RANKL (TNFSF11), a type II transmembrane protein, is a receptor activator of NF-κB (RANK) ligand. RANKL is an activator of RANK. When binding to RANK, it induces the differentiation of monocyte/macrophage-lineage cells into osteoclasts and leads to osteoclast precursor maturation. RANKL is critical for osteoclasts maturation, bone modeling, and bone remodeling, as well as the development of lymph nodes (LNs). RANKL/TNFSF11 Protein, Human (HEK293) is a recombinant human RANKL (G64-D245) without any tag, which is produced in HEK293 cell^[1][2].

RANKL (TNFSF11) belongs to TNF family. RANKL is a type II transmembrane protein and is a receptor activator of NF-κB (RANK) ligand. RANKL is an activator of NF-κB. RANKL binds to NF-κB and induces the differentiation of monocyte/macrophage-lineage cells into osteoclasts and leads to osteoclast precursor maturation. In bone tissue, RANKL is expressed by osteoblasts, osteocytes and immune cells, especially in osteoblasts and osteocytes^[1]. RANKL is also expressed by T cells and increases proliferation and survival of dendritic cells^[2].
Human RANKL shares 82.02% and 84.44% common aa identity with mouse and rat respectively. Human RANKL consists of cytoplasmic domain (1-47), helical domain (48-68), and extracellular domain (69-317). The soluble chain (140-317) is released when cleaved by enzymes such as matrix metalloproteinases (MMP3 or 7) and ADAM^[1][3].
RANKL is critical for osteoclasts maturation, bone modeling, and bone remodeling, as well as the development of lymph nodes (LNs)^[1].

RANKL (human, -5 ng/mL, 24 h) stimulates migration of a clear cell RCC cell line, Caki-1^[4].
RANKL (human, 24 h) stimulates PAa cell migration and invasion^[5].

RANKL (human, .4 or 2 mg/kg/day, s.c.) induces high bone turnover and decreases bone volume, density, and strength in C57BL/6J female mice^[6].

1.Immobilized human TNFSF11 at 2 μg/mL (100 μL/well) can bind human Osteoprotegerin-hFc and the EC₅₀ is 5-40 ng/mL.
2. Measured by its ability to induce TRAP activity, inducing osteoclast differentiation of RAW 264.7 mouse monocyte/macrophage cells. The ED₅₀ for this effect is < 20 ng/mL.
3. Loaded Narlumosbart (HY-P99966) on AHC2 biosensor, can bind RANKL/TNFSF11 Protein, Human (HEK293) with an affinity constant of <1.000E-11 M as determined in BLI assay.

• 
  Measured by its ability to induce osteoclast differentiation of RAW 264.7 mouse monocyte/macrophage cells. The ED₅₀ for this effect is 19.69 ng/mL, corresponding to a specific activity is 5.079×10⁴ units/mg.
• 
  Loaded Narlumosbart (HY-P99966) on AHC2 biosensor, can bind RANKL/TNFSF11 Protein, Human (HEK293) with an affinity constant of <1.000E-11 M as determined in BLI assay.

Human

HEK293

Tag Free

AAC51762.1 (G64-D245)

8600  [NCBI]

Partial

GSQHIRAEKAMVDGSWLDLAKRSKLEAQPFAHLTINATDIPSGSHKVSLSSWYHDRGWGKISNMTFSNGKLIVNQDGFYYLYANICFRHHETSGDLATEYLQLMVYVTKTSIKIPSSHTLMKGGSTKYWSGNSEFHFYSINVGGFFKLRSGEEISIEVSNPSLLDPDQDATYFGAFKVRDID

Approximately 27-33 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.

≥ 90%, as determined by reducing SDS-PAGE.

Lyophilized powder

1.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 5% trehalose, 5% mannitol, 0.01% Tween 80.
2.Lyophilized from a 0.22 μm filtered solution of 20 mM PB, 150 mM NaCl, pH 7.4.
Please refer to the lot-specific COA for specific buffer information.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.

• [1]. Ono T, et al. RANKL biology: bone metabolism, the immune system, and beyond. Inflamm Regen. 2020 Feb 7;40:2.  [Content Brief]

  [2]. Li B, et al. Roles of the RANKL-RANK Axis in Immunity-Implications for Pathogenesis and Treatment of Bone Metastasis. Front Immunol. 2022 Mar 21;13:824117.  [Content Brief]

  [3]. Tobeiha M, et al. RANKL/RANK/OPG Pathway: A Mechanism Involved in Exercise-Induced Bone Remodeling. Biomed Res Int. 2020 Feb 19;2020:6910312.  [Content Brief]

  [4]. Mikami S, et al. Increased RANKL expression is related to tumour migration and metastasis of renal cell carcinomas. J Pathol. 2009 Aug;218(4):530-9.  [Content Brief]

  [5]. Peng X, et al. Differential expression of the RANKL/RANK/OPG system is associated with bone metastasis in human non-small cell lung cancer. PLoS One. 2013;8(3):e58361.  [Content Brief]

  [6]. Lloyd SA, et al. Soluble RANKL induces high bone turnover and decreases bone volume, density, and strength in mice. Calcif Tissue Int. 2008 May;82(5):361-72.  [Content Brief]