Recombinant Mouse G-CSF

Recombinant Mouse G-CSF (Granulocyte Colony-Stimulating Factor) is widely used in research due to its critical role in regulating neutrophil production, mobilizing hematopoietic stem cells, and modulating immune responses in mouse models.

Key reasons to use recombinant mouse G-CSF in research applications include:

• Hematopoietic Stimulation: G-CSF is the principal cytokine controlling the production, survival, and differentiation of neutrophil precursors in the bone marrow, making it essential for studies on hematopoiesis and immune cell dynamics.
• Stem Cell Mobilization: G-CSF mobilizes hematopoietic stem and progenitor cells from the bone marrow into the peripheral blood, facilitating research on stem cell transplantation, engraftment, and recovery after myelosuppressive treatments.
• Innate Immunity and Infection Models: By increasing neutrophil counts and function, G-CSF enhances innate immune responses, making it valuable for infection, inflammation, and host defense studies in mice.
• Tissue Regeneration and Repair: G-CSF influences skeletal muscle development and regeneration by stimulating myoblast proliferation, and it has demonstrated neuroprotective and antiapoptotic effects in neuronal cells, broadening its utility to studies of tissue repair and neurobiology.
• Immunomodulation: G-CSF exerts immunosuppressive effects on monocytes, macrophages, dendritic cells, and T lymphocytes, allowing investigation into immune regulation and tolerance mechanisms.
• Chemotherapy and Transplantation Research: G-CSF is used to accelerate neutrophil recovery and reduce infection risk after chemotherapy or bone marrow transplantation, supporting preclinical studies of supportive care interventions.

Additional context:

• The receptor for G-CSF is expressed not only on hematopoietic cells but also on cardiomyocytes, skeletal muscle, and neurons, enabling research into its pleiotropic effects beyond hematopoiesis.
• Recombinant mouse G-CSF is species-specific, ensuring optimal activity in murine systems and avoiding cross-species variability that can occur with human G-CSF in mouse models.

In summary, recombinant mouse G-CSF is a versatile tool for studying hematopoiesis, immune responses, stem cell biology, tissue regeneration, and therapeutic interventions in mouse models, making it highly valuable for a wide range of biomedical research applications.

Yes, recombinant mouse G-CSF is widely used as a standard for quantification or calibration in ELISA assays designed to measure mouse G-CSF. This practice is supported by multiple technical sources and is standard in commercial ELISA kits.

Supporting details:

• Commercial ELISA kits for mouse G-CSF routinely use recombinant mouse G-CSF as the standard. These kits are validated to quantitate both recombinant and natural mouse G-CSF, with dose-response curves for natural G-CSF shown to be parallel to those obtained with recombinant standards.
• Quantification principle: The concentration of G-CSF in unknown samples is interpolated from a standard curve generated using recombinant mouse G-CSF. This allows for accurate calibration and quantification in serum, plasma, cell culture supernatants, and other biological fluids.
• Performance validation: ELISA kits using recombinant mouse G-CSF as a standard demonstrate high precision, recovery, and linearity across relevant sample types.
• Best practices: Ensure the recombinant protein is of high purity, properly formulated (e.g., in PBS), and free of interfering substances such as high endotoxin levels. Use freshly prepared standards and follow the kit or assay protocol for dilution and handling.

Additional considerations:

• Matrix effects: When quantifying G-CSF in complex biological samples, confirm that the recombinant standard behaves similarly to endogenous G-CSF in your assay matrix. Most validated kits report parallelism between recombinant and natural G-CSF curves, supporting reliable quantification.
• Bioactivity vs. quantification: Recombinant proteins labeled for ELISA standard use are not necessarily validated for bioassays. Use only proteins designated for ELISA calibration, not those intended for cell-based functional assays.
• Do not mix standards from different sources or kits: For consistent results, use standards and reagents from the same kit or validated source.

Summary of protocol:

1. Prepare serial dilutions of recombinant mouse G-CSF in the recommended diluent.
2. Run the ELISA according to the protocol, generating a standard curve.
3. Quantify unknown samples by interpolating their absorbance values against the standard curve.

In conclusion, recombinant mouse G-CSF is an appropriate and validated standard for ELISA quantification of mouse G-CSF, provided it is handled and prepared according to best practices and assay instructions.

Recombinant Mouse G-CSF has been validated for a broad range of scientific applications in published research, primarily related to hematopoiesis, immunology, cell biology, and regenerative medicine.

Key validated applications include:

• Functional Assays: Used to assess biological activity, such as stimulation of neutrophil production, survival, and differentiation from progenitor cells.
• Cell Culture: Supports proliferation and differentiation of hematopoietic stem cells, myeloid progenitors, and other responsive cell types.
• Western Blot: Detection and quantification of G-CSF or downstream signaling proteins in cell lysates.
• ELISA: Quantification of G-CSF levels or measurement of G-CSF-induced protein secretion in biological samples.
• Immunohistochemistry: Localization of G-CSF or its receptor in tissue sections.
• Stem/Immune Cell Maintenance or Differentiation: Used to maintain or differentiate stem cells and immune cells in vitro.
• Cell Adhesion Assays: Investigation of G-CSF’s effects on cell adhesion properties.

Published research applications:

• In vivo functional studies: Induction of emergency granulopoiesis and modulation of neutrophil and macrophage responses in mouse models of inflammation.
• Regenerative medicine: Promotion of muscle regeneration and protection against neuronal apoptosis in models of muscular dystrophy, spinal cord injury, ALS, and stroke.
• Cancer research: Investigation of G-CSF’s role in tumor microenvironment, myeloid-derived suppressor cell expansion, and immunosuppression.
• Hematopoietic stem cell mobilization: Enhancement of stem cell mobilization for transplantation and recovery after chemotherapy.
• Cardiac and neural studies: Stimulation of cardiomyocyte proliferation and neuroprotection in models of myocardial infarction and neurodegenerative diseases.

Summary Table of Validated Applications

These applications are supported by both product validation data and peer-reviewed studies, demonstrating the versatility of recombinant mouse G-CSF in basic and translational research.

To reconstitute and prepare Recombinant Mouse G-CSF protein for cell culture experiments, follow these general best practices based on manufacturer recommendations and scientific protocols:

Reconstitution:

1. Centrifuge the vial: Before opening, briefly centrifuge the lyophilized protein vial in a microcentrifuge (20–30 seconds) to ensure all powder is at the bottom.
2. Reconstitution buffer: Most recombinant mouse G-CSF proteins are reconstituted in sterile deionized water or sterile PBS. Some protocols recommend using a buffer containing 0.1% BSA (bovine serum albumin) to improve stability, especially for low-concentration solutions.
3. Concentration: Reconstitute to a concentration of 100 µg/mL (or as specified by the product datasheet). For example, add 100 µL of sterile water to 10 µg of protein.
4. Gentle mixing: Gently pipette up and down or swirl the vial to dissolve the protein. Avoid vigorous vortexing.
5. Incubation: Allow the solution to sit at room temperature for a few minutes to ensure complete dissolution.

Preparation for Cell Culture:

1. Further dilution: Dilute the reconstituted stock in cell culture medium or PBS as needed for your experiment. If preparing dilute working solutions, include 0.1–1% BSA or another carrier protein to prevent adsorption to plastic surfaces.
2. Sterile filtration: If necessary, filter-sterilize the solution using a 0.22 µm low-protein-binding filter.
3. Aliquoting: For long-term storage, aliquot the protein solution to avoid repeated freeze-thaw cycles.

Storage:

• Short-term: Store reconstituted protein at 2–8°C for up to one month.
• Long-term: For extended storage, aliquot and store at –20°C to –80°C. Avoid repeated freeze-thaw cycles.
• Do not refreeze thawed aliquots.

Additional Tips:

• Always refer to the product datasheet or certificate of analysis for specific instructions, as protocols may vary by manufacturer.
• Confirm protein integrity by running a small amount on SDS-PAGE if needed.
• For cell culture, ensure the final concentration of BSA or other additives is compatible with your cells.

By following these steps, you can ensure optimal activity and stability of recombinant mouse G-CSF for your cell culture experiments.