PE Rabbit anti-Human RANKL/CD254 Monoclonal Antibody for Flow Cytometry (Intracellular) - O14788

Core Product Specifications and Parameters

Biological Background: RANKL/CD254 Function and Localization

• TNFSF11, also known as RANKL, OPGL, ODF, TRANCE, and CD254, is a member of the tumor necrosis factor (TNF) ligand superfamily (UniProt: O14788).
• It functions as a key cytokine by binding to TNFRSF11A/RANK and TNFRSF11B/OPG, acting as an osteoclast differentiation and activation factor. Related references: PMID:22664871
• In the immune system, RANKL enhances dendritic cell-mediated stimulation of naive T-cell proliferation and regulates T-cell/dendritic cell interactions (By similarity).
• It induces osteoclastogenesis through activation of signaling pathways leading to NFATC1 nuclear translocation and osteoclast-specific gene transcription; also induces CREB1 activation and mitochondrial ROS via TMEM64-ATP2A2 axis (By similarity).
• Subcellularly, RANKL exists as a transmembrane protein on the cell membrane, but can also be shed and secreted, with detection in cytoplasm and secreted forms.
• Highest expression is found in peripheral lymph nodes, with weaker expression in spleen, peripheral blood leukocytes, bone marrow, heart, placenta, skeletal muscle, stomach, and thyroid.
• Post-translational modifications include glycosylation, and alternative splicing produces multiple isoforms, contributing to its functional diversity; mutations in TNFSF11 are associated with osteopetrosis.
• Keywords include cytokine, membrane, alternative splicing, developmental protein, glycoprotein, signal-anchor, and transmembrane helix.

Experimental Guidance and Technical Tips

• The immunogen spans the extracellular domain (aa 136-317), which may recognize both membrane-bound and soluble RANKL. For intracellular flow cytometry, ensure adequate cell permeabilization and consider including unstimulated controls.
• As a PE-conjugated antibody, protect from light and avoid repeated freeze-thaw cycles to maintain fluorescence intensity and antibody integrity.
• Validate performance in relevant cellular models such as activated T cells, osteoclast precursors, or RANKL-expressing cell lines. Titrate the antibody to determine the optimal staining concentration for your specific system.
• If using in combination with other fluorophores, carefully design compensation controls to avoid spectral overlap artifacts.

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