Anti-RANKL (CD254) Antibody, clone IK22-5

Tumor necrosis factor ligand superfamily member 11 (UniProt: O35235; also known as Osteoclast differentiation factor, ODF, Osteoprotegerin ligand, OPGL, Receptor activator of nuclear factor kappa-B ligand, RANKL, TNF-related activation-induced cytokine, TRANCE, CD254, RANKL) is encoded by the Tnfsf11 (also known as Opgl, Rankl, Trance) gene (Gene ID: 21943) in murine species. RANKL is a single-pass type II membrane protein that has a cytoplasmic domain (aa 1-48), a helical domain (aa 49-69) and an extracellular domain (aa 70-316). High levels of RANKL are detected in the peripheral lymph nodes and lower levels are detected in the spleen, peripheral blood Leukocytes, bone marrow, heart, placenta, skeletal muscle, stomach and thyroid. Its levels are shown to be up-regulated by T-cell receptor stimulation. RANKL has been identified to affect the immune system and control bone regeneration and remodeling. It is shown to augment the ability of dendritic cells to stimulate naive T-cell proliferation and is considered as an important regulator of interactions between T-cells and dendritic cells. It is shown to induce osteoclastogenesis by activating multiple signaling pathways in osteoclast precursor cells. In a murine model of collagen-induced arthritis, clone IK22-5 is shown to reduce bone loss at the site of inflammation. Also, it is reported to reduce joint destruction, but not prevent new bone formation, in an animal model of delayed-type hypersensitivity arthritis. (Ref.: Atkinson, SM., et al. (2016). Arthritis Res.Ther. 18; 28; Kamijo, s., et al. (2006). Biochem. Biophys. Res. Commun. 347(1); 124-132).

Recombinant fragment corresponding to the extracellular domain of murine RANKL (CD254).

Quality Control Testing

Evaluated by Flow Cytometry in L5178Y cells transfected with mTRANCE.

Flow Cytometry Analysis: A 1:250 dilution of this antibody detected RANKL in L5178Y cells transfected with mouse RANKL (mTRANCE).

Tested Applications

Inhibition Assay: A representative lot of this RANKL blocking antibody reduced the number of subcapsular sinus macrophages in wild-type mice, inhibited osteoclastogenesis, and inhibited erosive joint destruction. (Camara, A., et. al. (2019). Immunity. 50(6):1467-1481; Kamijo, S., et. al. (2006). Biochem Biophys Res. Commun. 347(1):124-32; Atkinson, S.M., et. al. (2016). Arthritis Res Ther. 18:28; Navet, B., et. al. (2018). Cancer (Basel). 10(11):398; Dougall, W.C., et. al. (2019). Clin Transl Immunology. 8(10):e01081).

Neutralizing: A representative lot of this antibody neutralized RANKL and reduced bone loss in collagen-induced arthritis and eliminated most Peyer s patches microfold cells (PP M cells). (Kamijo, S., et. al. (2006). Biochem Biophys Res Commun. 347(1):124-32; Knoop, K.A., et. al. (2009). J. Immunol. 183(9):5738-47).

Flow Cytometry Analysis: A representative lot detected RANKL (CD254) in Flow Cytometry applications ( Ahern, E., et. al. (2017). Clin Cancer Res. 23(19):5789-5801 ; Kamijo, S., et. al. (2006). Biochem Biophys Res Commun. 347(1):124-32).

Immunohistochemistry Applications: A representative lot detected RANKL (CD254) in Immunohistochemistry applications (Kamijo, S., et. al. (2006). Biochem Biophys Res Commun. 347(1):124-32).

Immunofluorescence Analysis: A representative lot detected RANKL (CD254) in Immunofluorescence applications (Knoop, K.A., et. al. (2009). J Immunol. 183(9):5738-47).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Anti-RANKL (CD254), clone IK22-5, Cat. No. MABF2802, is a rat monoclonal antibody that detects RANKL and is tested for use in Flow Cytometry, Immunofluorescence, Immunohistochemistry, Inhibition Assay, and Neutralizing.

Clone IK22-5 is a rat monoclonal antibody that detects murine Tumor necrosis factor ligand superfamily member 11 (RANKL/CD254). It targets an epitope within the extracellular domain.

Purified rat monoclonal antibody IgG2a in PBS without azide.

Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.