M CSF Human, Sf9 His
Description
Production and Formulation
M-CSF Human, Sf9 His is synthesized using baculovirus-infected Sf9 insect cells, leveraging their capacity for eukaryotic post-translational modifications. The production process involves:
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Expression: Secreted into the extracellular medium as a soluble protein .
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Purification: Proprietary chromatographic techniques, including immobilized metal affinity chromatography (IMAC) leveraging the His tag .
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Formulation: Provided as a 0.25 mg/mL solution in phosphate-buffered saline (pH 7.4) with 10% glycerol to prevent aggregation .
Biochemical Properties
| Property | Specification |
|---|---|
| Purity | >95% (SDS-PAGE, RP-HPLC) |
| Stability | 4°C for 2–4 weeks; -20°C for long-term storage |
| Endotoxin | <0.1 EU/µg (LAL method) |
| Bioactivity | ED₅₀ <3 ng/mL (M-NFS-60 cell assay) |
For long-term stability, adding carrier proteins (e.g., 0.1% HSA/BSA) is recommended to prevent adsorption losses .
Biological Activity and Mechanisms
M-CSF binds to the CSF-1 receptor (CSF-1R/CD115), triggering dimerization and activation of downstream pathways:
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MAPK/ERK and PI3K/AKT: Promote macrophage survival, proliferation, and differentiation .
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Osteoclast Regulation: Induces RANKL production, enhancing bone resorption .
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Immune Modulation: Upregulates phagocytosis, cytokine synthesis (e.g., IL-1, TNF-α), and tumor-associated macrophage activity .
In vitro studies demonstrate that M-CSF-derived macrophages (M(M-CSF)) exhibit distinct protease profiles compared to LPS/IFN-γ- or IL-4-polarized subtypes, with elevated MMP-9 and TIMP-2 levels .
Comparative Analysis of M-CSF Variants
Variants differ in glycosylation patterns, disulfide bonding, and species specificity. For example, murine M-CSF is inactive in humans, whereas human M-CSF cross-activates murine receptors .
Research Applications
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Macrophage Differentiation: Used to generate monocyte-derived macrophages for studying immune responses .
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Cancer Biology: Investigated for roles in tumor-associated macrophage polarization and VEGF-mediated angiogenesis .
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Bone Metabolism: Critical for osteoclast studies in osteoporosis and bone metastasis models .
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Structural Biology: Served as a template for crystallographic studies of CSF-1R binding .
Product Specs
Granulocyte/Macrophage Colony-Stimulating Factors (GM-CSFs) are cytokines that regulate hematopoiesis by influencing the production, differentiation, and function of granulocytes and monocytes-macrophages, two related white blood cell populations. Macrophage Colony-Stimulating Factor (MCSF) specifically stimulates cells of the monocyte/macrophage lineage. MCSF plays a crucial role in various biological processes, including immune responses, bone metabolism, lipoprotein clearance, fertility, and pregnancy.
Produced in Sf9 insect cells, MCSF is a single, glycosylated polypeptide chain consisting of 231 amino acids (specifically, amino acids 33 to 255). It possesses a molecular mass of 26.1 kDa. On SDS-PAGE analysis, the apparent molecular size will range from approximately 28 to 40 kDa. This MCSF protein is expressed with an 8-amino acid His tag at the C-terminus and purified using proprietary chromatographic techniques.
The product is a sterile, filtered solution that is colorless.
The MCSF protein solution is provided at a concentration of 0.25 mg/ml. It is formulated in Phosphate Buffered Saline (PBS) with a pH of 7.4 and contains 10% glycerol.
For short-term storage (2-4 weeks), the product can be stored at 4°C. For extended storage, it is recommended to freeze the product at -20°C. To further enhance long-term stability during storage, adding a carrier protein (either 0.1% HSA or BSA) is advisable. It is essential to avoid repeated freeze-thaw cycles to maintain product integrity.
The purity of the MCSF protein is determined by SDS-PAGE analysis and is greater than 95.0%.
The biological activity of MCSF is assessed through a cell proliferation assay using M-NFS-60 mouse myelogenous leukemia lymphoblast cells. The ED50 value, representing the effective concentration at which 50% of the maximal response is observed, is less than or equal to 3 ng/ml in this assay.
Macrophage colony-stimulating factor 1, CSF-1, M-CSF, MCSF, Lanimostim.
Sf9, Insect cells.
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Product Science Overview
M-CSF is primarily produced by monocytes, granulocytes, endothelial cells, and fibroblasts . It is a homodimeric glycoprotein growth factor that exerts its biological effects by signaling through a receptor tyrosine kinase known as CSF-1R or M-CSF-R, which is encoded by the c-fms proto-oncogene . The receptor shares structural features with other growth factor receptors, including the stem cell factor (SCF) receptor, platelet-derived growth factor receptor (PDGF-R), and Flt3/Flk-2 receptor tyrosine kinase .
M-CSF is involved in various biological processes, including:
- Regulation of Inflammatory Responses: It works in conjunction with other stimuli such as IFN-γ, LPS, and IL-4 to regulate inflammatory responses .
- Bone Resorption: M-CSF is essential for bone resorption by osteoclasts .
- Development and Regulation: It plays a role in the development and regulation of the placenta, mammary gland, and brain .
- Immunological Defense: M-CSF is crucial for the immunological defense mechanisms .
Recombinant human M-CSF, particularly the version expressed in Sf9 cells, is widely used in research and clinical applications. It is utilized in:
- Differentiation Studies: To study the differentiation of macrophages from peripheral blood monocytes .
- Cell Culture: For maintaining reproducible cell culture conditions without the need for time-consuming lot-to-lot testing .
- Functional Assays: In various functional assays to study the biological activity of M-CSF .
