Recombinant Mouse RANK

Recombinant Mouse RANK is used in research applications to study and manipulate key biological processes such as osteoclast differentiation, lymph node organogenesis, and immune system function. It provides a controlled, reproducible tool for dissecting the molecular mechanisms of the RANK-RANKL signaling axis in mouse models.

Essential context and supporting details:

• Osteoclast Differentiation and Bone Metabolism: RANK is a critical receptor for RANKL, and their interaction is essential for the differentiation of osteoclasts, the cells responsible for bone resorption. Recombinant Mouse RANK enables in vitro assays to study osteoclastogenesis, bone remodeling, and related pathologies such as osteoporosis and osteopetrosis.

• Immune System and Lymphoid Organ Development: RANK signaling is necessary for the formation of lymph nodes and the development of B cells in mice. Using recombinant RANK allows researchers to investigate immune cell development, lymphoid tissue formation, and the effects of RANK pathway modulation on immune responses.

• Functional Assays and Mechanistic Studies: Recombinant Mouse RANK is used in cell culture systems to activate or inhibit RANK signaling, facilitating studies on downstream pathways such as NF-κB, JNK, and AKT, which are involved in cell survival, differentiation, and immune regulation.

• Disease Modeling and Therapeutic Research: Manipulating RANK-RANKL interactions with recombinant proteins is fundamental in modeling diseases like osteoporosis, rheumatoid arthritis, and cancer-induced bone loss, as well as testing potential therapeutic interventions targeting this pathway.

Additional relevant information:

• Recombinant proteins offer high purity, batch-to-batch consistency, and defined activity, which are crucial for reproducible experimental results and mechanistic studies.
• Recombinant Mouse RANK can be used in combination with RANKL to dissect specific signaling events, validate genetic models, and perform bioassays for drug screening or pathway analysis.
• The use of recombinant proteins in mouse models is a standard approach for translating findings to human biology, given the conservation of the RANK-RANKL axis across species.

In summary, Recombinant Mouse RANK is indispensable for research focused on bone biology, immunology, and disease modeling due to its central role in osteoclast differentiation and lymphoid organ development, as well as its utility in controlled mechanistic studies and therapeutic research.

You can use recombinant Mouse RANK as a standard for quantification or calibration in ELISA assays, but only under specific conditions. The recombinant protein must be highly pure, well-characterized, and matched to the analyte detected by your assay. Additionally, it must be compatible with the antibodies and detection system used in your ELISA.

Key considerations:

• Intended Use: Recombinant proteins are commonly used as standards in ELISA assays, provided they are validated for this purpose. Many commercial ELISA kits use recombinant proteins as their standard curve calibrators.
• Protein Identity: Ensure that the recombinant Mouse RANK you have is the same form (e.g., full-length, extracellular domain, tag-free or tagged) as the native protein detected by your assay. Differences in isoform, post-translational modifications, or tags can affect antibody recognition and quantification accuracy.
• Validation: The recombinant standard should be validated for use in your specific ELISA format. This includes confirming that the standard curve generated with the recombinant protein is linear and covers the expected concentration range of your samples.
• Matrix Effects: If your samples are in a complex matrix (e.g., serum, plasma), matrix effects may cause differences in signal compared to the standard prepared in buffer. Internal controls or matrix-matched standards can help ensure accuracy.
• Documentation: Check the product datasheet or technical documentation for your recombinant Mouse RANK. It should state whether the protein is suitable for use as an ELISA standard. Some recombinant proteins are specifically formulated and tested for this application, while others are not.

Best Practices:

• Prepare a standard curve using serial dilutions of the recombinant Mouse RANK in the same buffer or matrix as your samples.
• Confirm that the standard curve is reproducible and covers the expected sample concentration range.
• If possible, compare the recombinant standard to a native protein standard or to a standard provided in a validated ELISA kit to ensure equivalence.

Caveats:

• Not all recombinant proteins are suitable as ELISA standards. Some are intended only for bioassays or functional studies and may not be recognized by the antibodies in your ELISA.
• If your ELISA kit is designed for RANKL (the ligand) and not RANK (the receptor), using recombinant RANK as a standard would not be appropriate, as the antibodies will not recognize the standard.

Summary Table:

In conclusion:
You may use recombinant Mouse RANK as a standard for ELISA quantification if it is validated for this purpose, matches the analyte detected by your assay, and is compatible with your assay conditions. Always consult the technical datasheet and, if possible, perform a validation experiment to confirm suitability in your specific ELISA system.

Recombinant Mouse RANK (TNFRSF11A) has been validated primarily for use in bioassays in published research, with applications focused on studying its biological activity in cell-based systems. Key validated applications include:

• Bioassays: Used to assess the activation of signaling pathways such as NF-κB and c-Jun N-terminal kinase, enhancement of T cell growth, dendritic cell function, and induction of osteoclastogenesis.
• Cellular Functional Studies: Employed to investigate the OPG/RANKL/RANK axis in inflammatory signaling, osteoclast differentiation, and lymph node organogenesis.
• Blocking Assays: Soluble recombinant RANK can be used to block RANKL (TRANCE)-induced biological activity, serving as a tool to dissect receptor-ligand interactions in vitro.
• ELISA Standard: Recombinant RANK is recommended as a standard in ELISA assays for quantifying RANK or RANKL activity in biological samples.
• Western Blot: Validated for use in Western blotting to detect RANK protein in cell lysates or tissue samples.
• SDS-PAGE: Used for protein characterization and purity assessment.

Supporting details:

• Published studies have used recombinant mouse RANK to explore its role in osteoclastogenesis, immune cell activation, and bone metabolism.
• The protein has been utilized in bioassays involving whole cells, particularly to study the effects of RANKL stimulation and the downstream signaling events.
• Recombinant RANK is also used as a blocking agent to inhibit RANKL-mediated effects, providing mechanistic insights into the RANK/RANKL/OPG pathway.

Additional relevant information:

• While recombinant RANK is less commonly used for direct cell culture or differentiation protocols compared to RANKL, its primary utility is in functional assays and mechanistic studies of receptor-ligand interactions.
• The protein is typically validated for use in mouse models and cell lines, with applications extending to immunology, bone biology, and inflammation research.

If you require details on specific published protocols or experimental setups, please specify the application area (e.g., osteoclastogenesis, immune modulation) for more targeted information.

To reconstitute and prepare Recombinant Mouse RANK protein for cell culture experiments, follow these standard steps to ensure protein stability and biological activity:

1. Centrifuge the Vial
   Briefly centrifuge the lyophilized protein vial before opening to collect all material at the bottom.

2. Reconstitution

   • Add sterile, endotoxin-free water or the buffer recommended in the product datasheet.
   • Typical reconstitution concentration is 0.1–1.0 mg/mL. For example, add 100–1000 µL of water to 0.1 mg of protein.
   • Gently mix by pipetting or swirling. Avoid vigorous shaking or vortexing to prevent protein denaturation or foaming.

3. Aliquoting and Storage

   • Divide the reconstituted protein into small aliquots to avoid repeated freeze-thaw cycles, which can degrade the protein.
   • Store aliquots at ≤ –20°C (not in a frost-free freezer) for long-term use.
   • For short-term use, aliquots can be kept at 2–8°C for up to a week, but always consult the specific product datasheet.

4. Working Dilutions

   • Prepare further dilutions in cell culture medium or buffer containing carrier protein (e.g., 0.1% BSA or FCS) to minimize adsorption and maintain stability.
   • Use low endotoxin solutions to avoid cell activation or toxicity.

5. Sterility

   • Ensure all solutions and tools are sterile. If sterility is uncertain, filter the final solution through a 0.22 µm sterile filter.

6. Application

   • The optimal working concentration for cell culture should be determined empirically, as it depends on the cell type and experimental design.
   • Add the protein directly to the culture medium at the desired final concentration.

Additional Notes:

• Always consult the specific product datasheet for any unique requirements, such as buffer composition or reconstitution instructions, as some recombinant proteins may require specific pH or additives for stability.
• Avoid repeated freeze-thaw cycles by aliquoting immediately after reconstitution.
• If the protein is sensitive to oxidation or aggregation, consider adding reducing agents or stabilizers as recommended by the manufacturer.

These steps are standard for most recombinant proteins used in cell culture, including RANK, unless otherwise specified in the product documentation.