Recombinant Mouse GM-CSF

Recombinant Mouse GM-CSF (Granulocyte-Macrophage Colony-Stimulating Factor) is a valuable tool for a wide range of research applications due to its critical roles in hematopoiesis, immune modulation, and inflammation. Here are key reasons why you should consider using recombinant mouse GM-CSF in your research:

1. Regulation of Myeloid Cell Development

• GM-CSF promotes the proliferation, differentiation, and survival of hematopoietic progenitor cells, leading to the generation of granulocytes, monocytes, macrophages, and dendritic cells.
• It is essential for the in vitro generation of dendritic cells (DCs) from bone marrow precursors, a common application in immunology research.

2. Immune System Activation and Modulation

• GM-CSF enhances the functional activity of mature myeloid cells, including antigen presentation, phagocytosis, oxidative metabolism, and cytokine production.
• It boosts host defense mechanisms against infections (bacterial, viral, fungal), making it useful in studies of infectious disease models.

3. Therapeutic and Disease Modeling Applications

• GM-CSF has been shown to improve survival and immune responses in mouse models of infection (e.g., viral, fungal), autoimmune diseases, and neurodegenerative conditions.
• It can be used to study the effects of immune modulation in models of cancer, inflammation, and autoimmune disorders.

4. Adjuvant and Vaccine Studies

• Recombinant mouse GM-CSF is frequently used as an adjuvant in DNA and protein vaccine studies to enhance immune responses and antigen presentation.

5. High Biological Activity and Reliability

• Recombinant forms are available with high specific activity (e.g., ED50 < 0.05 ng/ml), ensuring robust and reproducible results in bioassays and cell culture experiments.
• Carrier-free and endotoxin-free preparations are available for sensitive applications.

6. Versatile Experimental Use

• Suitable for a variety of applications, including:
  • Cell culture and differentiation assays
  • ELISA and Western blot standards
  • In vivo administration in mouse models
  • Functional studies of immune cell activation and survival

7. Relevance to Human Disease

• Insights gained from mouse GM-CSF studies often translate to understanding human immune responses and disease mechanisms, supporting translational research.

In summary, recombinant mouse GM-CSF is a powerful cytokine for studying immune cell biology, host defense, and therapeutic interventions, offering both versatility and biological relevance in research settings.

Yes, recombinant mouse GM-CSF is widely used as a standard for quantification or calibration in ELISA assays.

Recombinant mouse GM-CSF is specifically formulated and validated for use as an ELISA standard, allowing for accurate quantification of GM-CSF in biological samples. ELISA kits and protocols routinely use recombinant GM-CSF to generate standard curves, which are essential for determining the concentration of GM-CSF in unknown samples. The recombinant protein is typically reconstituted in phosphate-buffered saline (PBS) containing a carrier protein such as bovine serum albumin (BSA) at a concentration of at least 0.1% to ensure stability and reproducibility.

Key considerations for using recombinant mouse GM-CSF as an ELISA standard:

• Source and Purity: Recombinant GM-CSF produced in yeast or E. coli is suitable, provided it is highly purified (typically >95% by SDS-PAGE).
• Reconstitution: Use sterile PBS with 0.1–10% BSA or another appropriate carrier protein to prevent adsorption and loss of protein activity.
• Calibration: The standard curve generated with recombinant GM-CSF allows for quantification of both recombinant and natural GM-CSF in samples, as dose-response curves for natural and recombinant forms are parallel in validated assays.
• Bioactivity: The biological activity of the recombinant protein should be confirmed, typically via cell proliferation assays or other functional tests.
• Storage: After reconstitution, aliquot and store the standard at recommended temperatures (usually -20°C or below) to maintain stability.

Limitations and Best Practices:

• Ensure the recombinant GM-CSF standard matches the form used in your assay (e.g., glycosylation status, source organism) for optimal accuracy.
• Always follow the ELISA kit manufacturer’s instructions regarding standard preparation and dilution series.
• Use freshly prepared or properly stored aliquots to avoid degradation and variability.

In summary, recombinant mouse GM-CSF is an established and reliable standard for ELISA quantification, provided it is handled and prepared according to best laboratory practices.

Recombinant Mouse GM-CSF has been validated for a wide range of applications in published research, primarily in immunology, cell biology, and disease modeling. The most common validated applications include:

• Functional assays: Used to assess the biological activity of GM-CSF, such as stimulating proliferation, differentiation, and activation of myeloid lineage cells (e.g., dendritic cells, macrophages, granulocytes).
• Cell culture and differentiation: Widely used to generate bone marrow-derived dendritic cells (BMDCs) and macrophages from mouse progenitors in vitro, supporting studies of immune cell function and development.
• ELISA and Western blot: Employed as a standard or control in immunoassays to quantify or detect GM-CSF protein levels.
• Immunohistochemistry and immunoprecipitation: Used as a control or to validate antibody specificity in tissue and protein studies.
• Blocking and neutralization assays: Applied to study the functional role of GM-CSF by blocking its activity in vitro or in vivo.
• In vivo therapeutic and disease models:
  • Pulmonary and infectious disease models: Used to test therapeutic potential in murine models of pulmonary aspergillosis and influenza, where intranasal or systemic administration of recombinant GM-CSF improved host resistance and survival.
  • Neurodegenerative disease models: Evaluated for neuroprotective effects and immune modulation in models of Parkinson’s, Alzheimer’s, and Down syndrome.
  • Cancer immunotherapy: Used in studies where GM-CSF-transfected tumor cells serve as cancer vaccines or as an adjuvant to enhance immune responses.
  • Autoimmune and inflammatory disease models: Investigated in models of psoriasis and encephalitis to study its role in inflammation and immune regulation.
  • Hematopoietic recovery: Used to promote myeloid reconstitution after bone marrow transplantation or chemotherapy-induced neutropenia.

Additional validated uses:

• Adjuvant in vaccine studies: Co-administered with DNA vaccines to enhance immunogenicity.
• Bioassays: Used in a variety of published bioactivity assays to test immune cell responses, cytokine production, and cell survival.

Summary Table of Key Applications

These applications are supported by hundreds of peer-reviewed publications, demonstrating the versatility and reliability of recombinant mouse GM-CSF in both basic and translational research contexts.

Reconstitution Protocol

Reconstitute the lyophilized recombinant mouse GM-CSF protein by adding sterile, distilled water or sterile phosphate-buffered saline (PBS) to achieve a concentration of 0.1–1.0 mg/mL, with 100 μg/mL being a standard working concentration. Before opening the vial, centrifuge it briefly to bring the contents to the bottom. When reconstituting, gently pipette the solution down the sides of the vial rather than vortexing, as vigorous mixing can denature the protein.

Carrier Protein Addition

For enhanced stability, particularly during storage and dilution, add a carrier protein to your reconstitution solution. Bovine Serum Albumin (BSA) or human serum albumin (HSA) are the standard choices, with a minimum concentration of 0.1% in the final solution. If your experiments involve bovine-derived materials, consider using an alternative carrier protein such as equine serum albumin to avoid cross-reactivity. Alternatively, you can reconstitute the protein directly in complete cell culture medium containing serum, which provides both carrier protein and appropriate osmolarity for cell culture applications.

Storage and Stability

Store the lyophilized protein at ≤ –20°C, where it remains stable for 6–12 months when kept desiccated. Once reconstituted, follow these storage guidelines:

• Short-term storage (up to 1 month): Store at 2–8°C under sterile conditions
• Long-term storage (up to 3 months): Store at –20°C to –80°C

To prevent loss of potency, aliquot the reconstituted protein into working portions and avoid repeated freeze-thaw cycles by using a manual defrost freezer.

Preparation for Cell Culture

When preparing the protein for direct cell culture stimulation, you can reconstitute it in your complete cell culture medium rather than in PBS/BSA solution. This approach simplifies handling and ensures the protein is in the appropriate osmotic environment for your cells. Stock solutions can be prepared at 50–100 μg/mL in sterile buffer with 0.2–1% carrier protein, then further diluted as needed for your specific experimental requirements.

The recombinant mouse GM-CSF protein has a molecular weight of approximately 14 kDa and demonstrates high purity (>95%) with bioactivity typically showing an ED₅₀ of 5–30 pg/mL for stimulating myeloid cell proliferation.