Recombinant Human TRANCE/RANK L His Avi-tag Protein, CF

Recombinant Human TRANCE/RANK L/TNFSF11 (Catalog # AVI390) has a molecular weight (MW) of 85.7 kDa as analyzed by SEC-MALS, suggesting that this protein is a homotrimer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).

2 μg/lane of Biotinylated Recombinant Human TRANCE/TNFSF11/RANK L His-tag Avi-tag Protein (Catalog # AVI390) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 26-39 kDa.

When Recombinant Human RANK/TNFRSF11A Fc Chimera Protein (683-RK) is coated at 0.5 μg/mL (100 μL/well), the concentration of Biotinylated Recombinant Human TRANCE/TNFSF11/RANK L His-tag Avi-tag (Catalog # AVI390) that produces 50% optimal binding response is 3.00‑15.0 ng/mL.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human TRANCE/RANK L His Avi-tag Protein, CF Summary

• Additional Information: Analyzed by SEC-MALS. Biotinylated
• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human RANK/TNFRSF11A Fc Chimera Protein (Catalog # 683-RK) is coated at 0.5 μg/mL (100 μL/well), the concentration of Biotinylated Recombinant Human TRANCE/TNFSF11/RANK L His-tag Avi-tag that produces 50% optimal binding response is 3.00‑15.0 ng/mL
• Source: Chinese Hamster Ovary cell line, CHO-derived human TRANCE/TNFSF11/RANK L protein
  

  Avi-tag	HHHHHH	Human TRANCe/TNFSF11/RANK L (Ile140-Asp317) Accession # O14788.1
  N-terminus		C-terminus

• Accession #: O14788.1
• N-terminal Sequence:

  Gly of Avi-tag

• Structure / Form: Biotinylated via Avi-tag
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 24 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 26-39 kDa under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human TRANCE/RANK L His Avi-tag Protein, CF

• CD254 antigen
• CD254
• ODF
• OPGL
• OPGLOPTB2
• Osteoclast differentiation factor
• Osteoprotegerin ligand
• RANK L
• RANKL
• RANKLreceptor activator of nuclear factor kappa B ligand
• Receptor activator of nuclear factor kappa-B ligand
• sOdf
• TNF-related activation-induced cytokine
• TNFSF11
• TRANCE
• TRANCEODFhRANKL2
• tumor necrosis factor (ligand) superfamily, member 11
• tumor necrosis factor ligand superfamily member 11

Background

RANK L (receptor activator of NF-kappa B ligand), also called TRANCE (TNF-related activation-induced cytokines), OPGL (osteoprotegerin ligand), or ODF (osteoclast differentiation factor), is a 39‑45 kDa type II transmembrane (TM) protein in the tumor necrosis factor family, designated TNFSF11 (1‑5). RANK L, produced by osteoblasts and bone marrow stromal cells, is required for differentiation of osteoclasts and stimulates bone resorption (4, 6). It is also produced by activated T cells and augments dendritic cell stimulation; RANK L-/- mice lack lymph nodes and have impaired thymocyte development (1‑3, 6). The human RANK L cDNA encodes 317 amino acids (aa), including a 47 aa cytoplasmic domain, a 21 aa TM region, and a 249 aa extracellular domain (ECD) with two potential N‑linked glycosylation sites (note: Arg85‑Asp245 of Accession # AAC51762 is identical to Arg157‑Asp317 of SwissProt # O14788. This aa range contains the ECD trimerization and receptor‑binding motifs, but not ECD proteolytic cleavage sites). Within the ECD, human RANK L shares 89%, 89%, 93% and 95% aa identity with mouse, rat, bovine and porcine RANK L, respectively. Mouse RANK L can stimulate human osteoclast differentiation (4). Like most TNF family members, RANK L can form trimers (1). Soluble 31, 25 and 24 kDa forms of RANK L can be created by usage of alternate start sites at aa 74 or 146, or proteolytic cleavage by osteoblast- or stromal cell‑derived ADAM10 (after aa 139) or MMP14 (aa 146), or bone metastatic prostate tumor-derived MT1-MMP (aa 146) (5, 7, 8). Both TM and soluble extracellular RANK L act by engaging RANK receptors and are antagonized by the decoy receptor, OPG (osteoprotegrin) (2, 5). In resting cells, the majority of RANK L is stored in secretory lysosomes (9). In mammary epithelia, RANK L is upregulated by pregnancy hormones and is essential for the formation of a lactating mammary gland (10). In the brain, astrocyte RANK L mediates body temperature regulation (11). Pathologically, RANK L is thought to mediate post-menopausal osteoporosis, vascular calcification, progestin-induced breast cancer, cancer-induced bone disease, and osteopetrosis (in RANK L deficiencies) (12‑16). Our Avi-tag Biotinylated human TRANCE/RANK L features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

1. Leibbrandt, A. and J.M. Penninger (2008) Ann. N.Y. Acad. Sci. 1143:123.
2. Wong, B.R. et al. (1997) J. Biol. Chem. 272:25190.
3. Anderson, D.M. et al. (1997) Nature 390:175.
4. Lacey, D.L. et al. (1998) Cell 93:165.
5. Hikita, A. et al. (2006) J. Biol. Chem. 281:36846.
6. Kong, Y-Y. et al. (1999) Nature 397:315.
7. Accession # NP_143026 and EAX08679.
8. Sabbota, A.L. et al. (2010) Cancer Res. 70:5558.
9. Aoki, S. et al. (2010) J. Bone Miner. Res. 25:1907.
10. Fata, J.E. et al. (2000) Cell 103:41.
11. Hanada, R. et al. (2009) Nature 426:505.
12. Osako, M.K. et al. (2010) Circ. Res. 107:466.
13. Schramek, D. et al. (2010) Nature 468:98.
14. Gonzalez-Suarez, E. et al. (2010) Nature 468:103.
15. Dougall, W.C. and M. Chaisson (2006) Cancer Metastasis Rev. 25:541.
16. Sobacchi, C. et al. (2007) Nat. Genet. 39:960.

Publications for TRANCE/TNFSF11/RANK L (AVI390)(1)

Publication using AVI390

• H Qiu, C Hosking, E Rothzerg, A Samantha, K Chen, V Kuek, H Jin, S Zhu, A Vrielink, K Lim, M Foley, J Xu ADR3, a next generation i-body to human RANKL, inhibits osteoclast formation and bone resorption The Journal of Biological Chemistry, 2023-01-09;0(0):102889. 2023-01-09 [PMID: 36634847] (Bioassay, Human)

Bioinformatics

• Uniprot:
  • O14788.1()