Recombinant Human M-CSF

Recombinant Human M-CSF (Macrophage Colony-Stimulating Factor) is widely used in research applications to promote the survival, proliferation, and differentiation of monocytes and macrophages, making it essential for studies involving hematopoiesis, immune response, and tissue repair.

Key scientific reasons to use recombinant human M-CSF include:

• Macrophage and Monocyte Differentiation: M-CSF is critical for driving the differentiation of hematopoietic stem cells into monocytes and macrophages in vitro, enabling the generation of these cell types for functional assays, cell culture, and differentiation studies.

• Cell Survival and Proliferation: It supports the survival and proliferation of peripheral blood monocytes and macrophages, which is necessary for experiments requiring large numbers of these cells.

• Functional Activation: M-CSF enhances key monocyte and macrophage functions, such as phagocytosis, microbial killing, cytokine production (e.g., IL-1, TNFα, IFNγ), and antigen presentation, making it valuable for immunological and infection models.

• Tissue Repair and Regeneration: M-CSF plays a role in tissue repair and regeneration by promoting macrophage-mediated processes, which is relevant for studies on wound healing, fibrosis, and regenerative medicine.

• Disease Modeling and Therapeutic Research: It is used in models of infection, transplantation, cancer, and inflammatory diseases to study immune cell dynamics, host defense mechanisms, and therapeutic interventions.

• Macrophage Polarization: M-CSF can polarize macrophages toward the M2 phenotype, which is associated with anti-inflammatory and tissue-repair functions, useful for research into immune modulation and chronic inflammation.

• Osteoclast Biology: M-CSF is a potent stimulator of osteoclast differentiation and activity, making it essential for bone biology and osteoporosis research.

• GMP-Grade Applications: Recombinant human M-CSF is available in GMP-grade for translational research and cellular therapy development, such as manufacturing phagocytic macrophages for therapeutic use.

Typical applications include:

• Differentiation and maturation of monocytes/macrophages from progenitor cells
• Survival and apoptosis assays
• Functional assays (phagocytosis, cytokine release, cytotoxicity)
• Infection and transplantation models
• Tissue repair and regeneration studies
• Osteoclastogenesis assays

In summary, recombinant human M-CSF is indispensable for any research requiring reliable generation, maintenance, and functional activation of human monocytes and macrophages, as well as for modeling immune responses, tissue repair, and disease mechanisms in vitro and in vivo.

Yes, recombinant human M-CSF can be used as a standard for quantification or calibration in ELISA assays, provided it is properly characterized and compatible with the detection antibodies in your specific ELISA kit.

Several sources confirm this practice:

• R&D Systems recommends using their recombinant human M-CSF (Catalog # 216-MC) for use as an ELISA standard, especially when formulated with BSA for stability and compatibility in immunoassays. The protein is well-characterized and suitable for generating standard curves in quantitative ELISA applications.
• BioLegend notes that their recombinant human M-CSF is suitable as an ELISA standard, but cautions that it is not tested for bioassay applications (i.e., functional cell-based assays), reinforcing its intended use for immunoassay calibration.
• Abcam’s SimpleStep ELISA Kit (ab245714) is calibrated against a highly purified human M-CSF standard, and the kit documentation confirms that recombinant M-CSF is recognized by the assay, with no observed cross-reactivity or interference.

Key Considerations:

• Ensure the recombinant M-CSF is highly purified and properly quantified (e.g., by amino acid analysis or other validated methods).
• Confirm that the epitopes recognized by your ELISA kit’s capture and detection antibodies are present on the recombinant protein.
• For best results, use the recombinant protein in the same matrix as your samples (e.g., diluted in assay buffer or serum/plasma) to minimize matrix effects.

In summary, recombinant human M-CSF is widely used and accepted as a standard for ELISA quantification, as long as it is compatible with your assay system and appropriately validated.

Recombinant Human M-CSF has been validated for a broad range of applications in published research, primarily involving the regulation, proliferation, differentiation, and survival of monocytes, macrophages, osteoclasts, and related progenitor cells. The most commonly validated applications include:

• Cell culture and differentiation assays: Used to differentiate monocytes into macrophages or osteoclasts, including from peripheral blood monocytes and CD14+ monocytes.
• Functional assays and bioassays: Validated for promoting proliferation of cell lines such as M-NFS-60, and for assessing survival, apoptosis, and functional activation of monocytes/macrophages.
• ELISA and immunoassays: Used as a standard or control in ELISA for cytokine quantification and validation.
• Western blot and SDS-PAGE: Used as a positive control or for protein validation in immunoblotting and electrophoresis.
• Immunohistochemistry: Applied as a control or for validating antibody specificity in tissue sections.
• Hematopoietic stem and progenitor cell research: Supports expansion and differentiation protocols for hematopoietic and mesodermal lineages.
• Osteoclastogenesis and bone biology studies: Induces RANKL production and stimulates osteoclast differentiation and bone resorption activity.
• Macrophage polarization and immune modulation: Used to study macrophage activation, cytokine production, and polarization in response to various stimuli.
• Tumor biology and angiogenesis research: Investigates the role of M-CSF in tumor-associated macrophage function, VEGF induction, and cancer prognosis.
• Neuroscience and microglia studies: Supports differentiation of microglia from iPSCs or other progenitors for neuroinflammation and neurodegeneration models.

Representative published research applications include:

• Differentiation of human iPSC-derived microglia and macrophages.
• Functional bioassays measuring proliferation of M-NFS-60 cells.
• Studies on macrophage-mediated immune responses, phagocytosis, and cytokine release.
• Osteoclast differentiation and bone resorption assays.
• Investigation of M-CSF’s role in inflammatory diseases, atherosclerosis, and cancer.

Summary Table: Validated Applications for Recombinant Human M-CSF

These applications are supported by both product validation data and peer-reviewed research, demonstrating the versatility of recombinant human M-CSF in immunology, stem cell biology, bone research, and disease modeling.

To reconstitute and prepare Recombinant Human M-CSF protein for cell culture experiments, follow these best-practice steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized powder is at the bottom.
• Reconstitute the protein in an appropriate sterile buffer. Common options include:
  • Sterile water to a concentration of 0.1–1.0 mg/mL.
  • Sterile PBS (pH 7.2–7.4), optionally containing 0.1–1% carrier protein such as bovine serum albumin (BSA) or human serum albumin (HSA), especially if the protein will be stored or used at low concentrations.
  • Some protocols recommend 20 mM Tris-HCl, pH 8.0 as an alternative buffer.
• Mix gently by swirling or tapping; do not vortex, as this can denature the protein.
• Allow the protein to fully dissolve at room temperature for 10–30 minutes, with occasional gentle mixing.
• Optional: If not using immediately, dilute the reconstituted protein with buffer containing 0.1% BSA or HSA to stabilize and prevent adsorption to plastic.
• Prepare working aliquots to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at −20°C to −80°C for long-term storage; short-term storage (up to 1 week) at 4°C is possible if carrier protein is present.

Typical working concentrations for cell culture are in the range of 0.5–100 ng/mL, but the optimal concentration should be determined empirically for your specific application.

Summary Table: Reconstitution Options

Key technical notes:

• Always check the product-specific datasheet for any unique requirements.
• Avoid reconstituting at concentrations above 1 mg/mL to prevent solubility issues.
• Use low-protein binding tubes for reconstitution and storage.
• Confirm protein integrity by SDS-PAGE if needed.

These guidelines ensure optimal activity and stability of recombinant M-CSF for cell culture applications.