Anti-Mouse RANKL (CD254) – Purified in vivo PLATINUM™ Functional Grade

Common in vivo applications of clone IK22/5 in mice primarily involve functional blockade of RANKL (Receptor Activator of Nuclear factor Kappa-Β Ligand), a key regulator of bone metabolism, immune cell interactions, and tissue development. The antibody is widely used to probe and manipulate the RANKL pathway in various physiological and disease contexts.

Key in vivo research applications include:

• RANKL Blockade: IK22/5 is commonly administered in vivo to neutralize mouse RANKL activity, preventing its binding to the RANK receptor and thereby inhibiting RANKL-mediated signaling.
• Bone Resorption/Osteoclastogenesis Studies: Given RANKL’s essential role in osteoclast differentiation and function, IK22/5 is used to study bone loss, osteoporosis, arthritis, and cancer metastases to bone by blocking osteoclast formation and activity in mouse models.
• Immune Regulation: RANKL and RANK are involved in T cell and dendritic cell interactions, T cell activation, and lymph node development—IK22/5 can be used to dissect these immune processes in vivo.
• Developmental Biology: The antibody can be applied to investigate RANKL’s role in thymic microenvironment formation, mammary gland development, and lymph node organogenesis in genetic or pharmacological mouse models.
• Disease Models: IK22/5 is used in models of autoimmune disease, inflammation, and cancer to study therapeutic blockade of RANKL and downstream effects on bone, immune, or tissue homeostasis.

Additionally, clone IK22/5 is validated for other applications such as flow cytometry for detection of RANKL-expressing cells and immunohistochemistry for tissue localization, but in vivo use is centered on functional inhibition of RANKL signaling.

In summary, IK22/5 is a standard tool for in vivo neutralization of RANKL to study bone metabolism and immune modulation, representing a key reagent in mouse models of osteoimmunology, development, and disease.

Commonly used antibodies or proteins studied with IK22/5 (an anti-mouse RANKL antibody) in the literature include those targeting key immune checkpoint molecules (such as PD-1, PD-L1, and sometimes CTLA-4), as well as markers relevant to osteoblast and osteoclast biology (such as RANK and bone lineage markers).

Key examples include:

• Anti–PD-1: Frequently combined with IK22/5 to study synergistic effects in tumor and metastasis models, particularly in immuno-oncology research.
• Anti–PD-L1: Used in similar combination therapy contexts with IK22/5.
• Anti–CTLA-4: Sometimes included to interrogate additional immune regulatory pathways.
• Anti–RANK (e.g., clone LOB14-8): Used alongside IK22/5 to investigate the RANK/RANKL axis, especially in bone and immune cell differentiation studies.
• Osteoblast and osteoclast markers: Proteins or staining assays such as CFU-F for osteoblast progenitors, Von Kossa for mineralized nodules, and TRAP staining for osteoclasts are used to assess effects on bone cell populations in combination with IK22/5.
• Flow cytometry markers: For multiparametric analysis, IK22/5 may be used with lineage markers like B220 (B cells), GL7 (germinal center B cells), and Thy1.2 (T cells).

In summary, researchers commonly pair IK22/5 with antibodies against PD-1, PD-L1, CTLA-4, RANK, and various bone lineage and immune cell markers in studies ranging from tumor models to bone metabolism.

Clone IK22/5 is a rat monoclonal antibody directed against mouse RANKL (CD254, also known as TRANCE). Key findings from scientific literature citing this clone highlight its critical role in osteoporosis, cancer metastasis, and immune regulation studies.

Principal findings from IK22/5 citations:

• Potent RANKL Neutralization: IK22/5 is widely confirmed to completely neutralize mouse RANKL in vitro and in vivo, effectively inhibiting osteoclast differentiation and bone resorption by reducing expression of osteoclast markers (TRAP, CTSK) and suppressing formation of TRAP-positive multinucleated cells.
• Impact on Bone Remodeling and Disease Models: IK22/5 administration in mice robustly blocks RANKL activity, resulting in increased bone mass, impaired osteoclastogenesis, and resistance to experimental bone loss. Its use has enabled detailed studies of bone homeostasis, osteopetrosis, and anabolic/parathyroid hormone effects in the absence of osteoclasts. Strain differences and synergism with agents like zoledronic acid have also been reported.
• Cancer Research: Blockade of RANKL with IK22/5 significantly restricts bone metastatic growth in models of prostate cancer and melanoma. Combination therapy with anti-RANKL and anti-PD-1 antibodies yields superior control of metastatic burden compared to single checkpoint blockade, supporting an immunoregulatory role for RANKL in tumor microenvironments.
• Immunological Functions: Antibody targeting with IK22/5 has shown RANKL’s importance in dendritic cell survival, regulation of T cell-dependent responses, and lymphocyte differentiation. Knockout or antibody blockade studies have demonstrated defective immune organ structure and lymphocyte development in vivo.
• Experimental Applications: IK22/5 is extensively used for flow cytometry, immunohistochemistry, and in vivo functional neutralization to dissect RANKL biology.

Technical notes from literature and suppliers:

• IK22/5 is recommended for use in flow cytometry at ≤0.5 µg/test, and for in vivo studies at 1 mg/kg repeated dosing or specified regimens, depending on disease model.
• The antibody specifically recognizes mouse (not human) RANKL and is available in various conjugated forms for research use.

Summary Table: Key Applications of IK22/5

These findings collectively have established IK22/5 as a gold standard tool in mouse RANKL research and a pivotal reagent for studying the RANK/RANKL/OPG signaling pathway in bone and immune systems.

Dosing regimens of clone IK22/5, a rat anti-mouse RANKL monoclonal antibody, vary substantially depending on the experimental context and mouse model, with typical in vivo doses ranging from 0.5 mg/kg to 2.5 mg/kg per injection, commonly administered via intraperitoneal or subcutaneous routes at intervals from a single dose to weekly dosing.

Supporting details and context:

• Cancer models: In studies targeting tumor-bearing mice, IK22/5 was often given as a single intraperitoneal dose of 1 mg/kg.
• Bone disease or developmental studies:
  • In a study on early-life treatment, 1-week-old mice received a single subcutaneous injection of 2.5 mg/kg anti-RANKL (clone IK22/5). In this context, the main outcome measured was the effect on bone growth and tooth eruption, as well as long-term skeletal effects.
  • Other studies assessing long-term bone turnover and pathological bone growth administered the antibody on a weekly basis at 2.5 mg/kg for several weeks.
  • For fibrous dysplasia models, mice received repeat injections following specific monitoring protocols, but the reported regimens tend to mirror the therapeutic approaches above, with most injected at 1–2.5 mg/kg intervals.
• Immunology or tumor immunology models: For immune checkpoint/bispecific antibody experiments, 100 μg per injection (approximately 4–5 mg/kg in a 20–25 g mouse) was used on days −1, 0, and 2 relative to tumor cell inoculation.
• In vitro/Ex vivo experiments: For cell culture studies examining osteoclastogenesis, IK22/5 is used at 0–1000 nM for 3–5 days, titrated for dose dependency.

Regimen determination factors:

• Purpose of study: Blockade for bone disease often uses higher, repeat doses; immunological blockade may need lower, shorter-duration regimens.
• Mouse model specifics: Age, weight, disease model, and genetic background impact the optimal dosing schedule.
• Antibody lots and source: Some suppliers recommend a pilot titration for optimal results, and in flow cytometry, suggested use is ≤0.5 μg per test.

Summary Table: Common IK22/5 Dosing Regimens in Mice

Note: Individual studies may deviate from these regimens, so consulting the primary literature or supplier recommendations for your specific application is essential. Dosing should be carefully titrated for each experimental context.