Recombinant Mouse Oncostatin M

Recombinant Mouse Oncostatin M (OSM) is a valuable tool for a wide range of research applications due to its well-characterized biological activities and its role in multiple physiological and pathological processes. Here are several key reasons why you should consider using Recombinant Mouse Oncostatin M in your research:

1. Well-Defined Biological Functions

Recombinant Mouse OSM is a pleiotropic cytokine belonging to the interleukin-6 (IL-6) family. It plays critical roles in:

• Inflammation: OSM is a potent pro-inflammatory cytokine involved in the regulation of immune responses, including the activation of signaling pathways such as JAK/STAT, MAPK, and PI3K/AKT in various cell types.
• Bone Metabolism: OSM promotes both osteoblast differentiation and osteoclast formation, making it essential for studies on bone development, remodeling, and diseases such as osteoporosis and arthritis.
• Wound Healing: OSM accelerates wound healing by stimulating collagen and glycosaminoglycan production in fibroblasts, which is relevant for research in tissue repair and regenerative medicine.
• Cell Cycle Regulation: OSM can regulate the cell cycle by modulating cyclin and CDK expression, influencing cell proliferation and differentiation in various cell lines.

2. High Purity and Bioactivity

Recombinant Mouse OSM is produced in E. coli and is available in high purity (≥ 99%). Its bioactivity is confirmed by its ability to induce OSM/OSM receptor signaling, such as STAT3 phosphorylation, in target cells. This ensures reliable and reproducible results in your experiments.

3. Versatile Applications

Recombinant Mouse OSM can be used in a variety of experimental settings, including:

• In vitro studies: To investigate the effects of OSM on cell proliferation, differentiation, and signaling pathways in different cell types.
• In vivo studies: To study the role of OSM in animal models of inflammation, bone disease, and wound healing.
• Drug Screening and Development: OSM can be used in NMR-based small molecule screening and structure-based drug design to identify inhibitors or modulators of OSM signaling.

4. Relevance to Disease Models

Elevated levels of OSM have been found in various diseases, including:

• Rheumatoid Arthritis (RA): OSM is involved in the pathogenesis of RA by promoting the secretion of pro-inflammatory cytokines and enhancing the expression of OSMR and IL-1β receptors in fibroblast-like synoviocytes (FLS).
• Inflammatory Bowel Disease (IBD): OSM expression is increased in colonic biopsies of patients with active IBD, making it a relevant target for studying intestinal inflammation.
• Cancer: OSM has been implicated in cancer progression and metastasis, particularly in breast cancer, where it promotes the expression of proteases and proangiogenic proteins.

5. Support for Advanced Research Techniques

Recombinant Mouse OSM is suitable for advanced research techniques such as:

• NMR-based studies: Isotopically enriched OSM can be used for NMR titrations and structure-based drug design.
• ELISA and Immunoblotting: OSM can be used to detect and quantify phosphorylation of STAT3 and other signaling molecules.

6. Consistency and Reproducibility

Using a recombinant form of OSM ensures consistency and reproducibility across experiments, which is crucial for generating reliable data and drawing meaningful conclusions.

Summary

Recombinant Mouse Oncostatin M is a powerful and versatile tool for researchers studying inflammation, bone metabolism, wound healing, and cancer. Its high purity, bioactivity, and relevance to various disease models make it an excellent choice for both basic and applied research applications.

Recombinant Mouse Oncostatin M can be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose and matches the assay requirements.

Key considerations and supporting details:

• ELISA standards are typically recombinant proteins: Commercial ELISA kits for mouse Oncostatin M (OSM) use E. coli-expressed recombinant mouse OSM as the standard for generating calibration curves. These standards are specifically validated for quantification in the context of the kit's antibodies and assay conditions.

• Validation is essential: Not all recombinant OSM preparations are suitable as ELISA standards. The recombinant protein must be of high purity, correctly folded, and biologically active. It should also be compatible with the antibodies used in your ELISA (i.e., recognized equivalently to native OSM). Some recombinant proteins are labeled for use in bioassays but not as ELISA standards, and vice versa.

• Parallelism and recovery: The ELISA kit documentation demonstrates that recombinant mouse OSM produces dose-response curves parallel to those of natural OSM, indicating it is suitable for quantification. Recovery experiments confirm that recombinant OSM can be accurately measured in various sample matrices.

• Carrier-free vs. carrier-added: For ELISA standards, carrier-free recombinant proteins are generally preferred to avoid interference from carrier proteins such as BSA, unless the assay specifically requires a carrier.

• Practical protocol: To use recombinant mouse OSM as a standard:

  • Prepare a serial dilution of the recombinant protein in the same buffer or matrix as your samples.
  • Generate a standard curve in each assay run.
  • Ensure the recombinant protein is stored and handled according to manufacturer or best practice guidelines to maintain stability and activity.

• Check your ELISA kit requirements: If you are using a commercial ELISA kit, it is best to use the standard provided or one that is specifically validated for that kit. If developing your own assay, ensure your recombinant OSM is validated for use as a standard with your antibody pair and detection system.

In summary: You can use recombinant mouse Oncostatin M as a standard for ELISA quantification if it is validated for this application, is of high purity, and is recognized by your assay antibodies. Always confirm compatibility with your specific assay system and, if possible, compare the standard curve generated with recombinant OSM to that of a known standard or native protein for accuracy.

Recombinant Mouse Oncostatin M (OSM) has been validated in published research for a range of applications, primarily in cell-based assays, in vivo studies, and protein detection methods.

Key validated applications include:

• Bioassays (cell-based functional assays):

  • Induction of lipolysis and suppression of insulin response in 3T3-L1 adipocytes.
  • Stimulation of mouse synovial fibroblasts and investigation of cytokine responses in joint connective tissue cells.
  • Promotion of tumor progression and metastasis in cancer models, including skin squamous-cell carcinoma and breast cancer.
  • Investigation of endothelial activation and inflammatory responses in vascular and adipose tissues.
  • Enhancement of mast cell growth via fibroblast-dependent pathways.
  • Modulation of immune cell trafficking, such as increasing CCL21 expression in endothelial cells to facilitate dendritic cell migration.
  • Promotion of neurogenic heterotopic ossification and nerve fiber elongation in skin.

• In vivo assays:

  • Local administration in mouse models to study joint inflammation, tumor progression, and nerve elongation.
  • Investigation of OSM’s role in liver engraftment and regeneration using pluripotent stem cell-derived hepatocytes.
  • Studies of OSM’s effects on retinal regeneration and vascular modulation in ocular disease models.

• Protein detection and signaling studies:

  • Western blot analysis to assess OSM-induced signaling events, such as STAT3 activation and osteocyte-like differentiation.
  • ELISA standardization and cytokine quantification in cell culture supernatants.

• Other validated uses:

  • Regulation of cytokine production (e.g., IL-6, G-CSF, GM-CSF) from endothelial cells.
  • Studies of OSM’s effects on bone, muscle, and heart regeneration, as well as its role in intestinal inflammation via the OSMR pathway.

Summary Table of Validated Applications

Additional Notes:

• Most studies use recombinant mouse OSM to probe its biological activity in mouse-derived cells or in mouse models, focusing on inflammation, cancer, metabolism, tissue regeneration, and immune cell function.
• The protein is also used to dissect OSM/OSMR signaling mechanisms and to evaluate therapeutic strategies targeting this pathway in preclinical models.

If you need protocols or more specific details for a particular application, please specify the context or experimental system.

To reconstitute and prepare Recombinant Mouse Oncostatin M (OSM) protein for cell culture experiments, dissolve the lyophilized protein in sterile water or PBS at a concentration between 0.1–0.5 mg/mL, then dilute as needed for your assay.

Essential protocol steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Reconstitution: Add sterile distilled water or sterile PBS to achieve a final concentration of 0.1–0.5 mg/mL. Some protocols recommend 0.1 mg/mL in sterile water, while others suggest 25–100 μg/mL in sterile PBS.
• Mix gently: Avoid vortexing or vigorous pipetting to prevent protein denaturation.
• Carrier protein/stabilizer: For long-term storage or to prevent adsorption, add a carrier protein such as 0.1% BSA, 5% HSA, 10% FBS, or 5% trehalose.
• Aliquot: Divide the reconstituted solution into small aliquots to minimize freeze-thaw cycles.
• Storage: Store aliquots at –20°C or below for long-term use. For short-term use (up to one week), store at 2–8°C.
• Dilution for cell culture: Dilute the stock solution in cell culture medium or buffer immediately before use to the desired working concentration, typically in the range of ng/mL to low μg/mL, depending on your experimental design.

Additional notes:

• Always consult the specific product datasheet for recommended buffer and concentration, as formulations may vary.
• Avoid foaming and bubbles during reconstitution, as these can denature the protein.
• If using PBS, ensure the pH is 7.2–7.4 for optimal protein stability.
• For bioassays, confirm the final buffer is compatible with your cell type and experimental conditions.

Summary Table: Reconstitution Options

These steps will ensure the recombinant Mouse OSM protein is properly prepared for reliable cell culture experiments.