Anti-Mouse CSF1 – Purified in vivo PLATINUM™ Functional Grade

Common In Vivo Applications of Clone 5A1 in Mice

Clone 5A1 is a monoclonal antibody that targets mouse Colony Stimulating Factor 1 (CSF-1, also known as macrophage colony-stimulating factor, M-CSF). Its ability to neutralize CSF-1 in vivo has made it a valuable tool in mouse models for studying the biology of monocytes, macrophages, and related immune responses.

Key Applications

• Macrophage Depletion Studies
  The primary in vivo application of clone 5A1 is to deplete or suppress macrophage populations by neutralizing CSF-1, which is essential for the survival, proliferation, and differentiation of monocytes and macrophages. This approach is widely used to investigate the role of macrophages in disease models, including cancer, inflammation, and infectious diseases.
• Atherosclerosis Research
  Because CSF-1 produced in vessel walls is implicated in the development and progression of atherosclerosis, clone 5A1 has been used to investigate the contribution of macrophages to plaque formation and stability in mouse models of cardiovascular disease.
• Bone Marrow and Hematopoiesis Studies
  Clone 5A1 inhibits the colony formation of bone marrow progenitor cells, making it useful for studying the regulation of hematopoiesis and the differentiation of myeloid lineage cells in vivo.
• Functional Assays of Biological Pathways
  The antibody is employed in bioanalytical assays to study pathways affected by CSF-1 signaling, including those involved in immune regulation and tissue homeostasis.

Mechanism of Action

Clone 5A1 binds to mouse CSF-1 and neutralizes its biological activity, preventing it from interacting with its receptor (CSF-1R) on target cells such as macrophages and progenitor cells. This blockade leads to reduced survival and proliferation of CSF-1-dependent cell populations in vivo.

Technical Specifications

• Isotype: Rat IgG1, κ
• Formulation: Typically supplied in PBS, suitable for direct in vivo injection
• Purity and Safety: High purity (≥95%), low endotoxin, and animal-free production ensure suitability for in vivo use
• Storage: Recommended at 4°C; freezing is not advised

Summary Table

Conclusion

Clone 5A1 is most commonly used in vivo in mice to deplete or modulate macrophage populations by neutralizing CSF-1, enabling researchers to dissect the roles of these cells in health, disease, and tissue homeostasis. Its applications span immunology, cancer research, cardiovascular disease, and hematopoiesis.

Commonly used antibodies or proteins paired with 5A1 (typically referencing the monoclonal antibody clone 5A1 against ABCA1) in the literature include those to validate or visualize different cellular compartments, cholesterol metabolism, or other transporter proteins and cell-type markers. Some of the most cited proteins and antibodies are:

• ABCA1 (target of 5A1): 5A1 is widely recognized as an antibody against ABCA1, a crucial transporter in cholesterol efflux and lipid metabolism studies.

Frequently co-used antibodies or proteins in ABCA1 research include:

• ATP5A1 (ATP synthase, F1 complex, alpha subunit): Used as a mitochondrial marker or loading control in Western blotting, and in studies monitoring energetic and metabolic changes associated with cholesterol transport.
• COL5A1 (Collagen, type V, alpha 1): Used in tissue integrity, extracellular matrix studies, and co-localization studies—particularly in contexts where cholesterol efflux affects extracellular composition.
• NR5A1 (Nuclear receptor subfamily 5, group A, member 1, SF-1): Relevant when connecting cholesterol handling to steroidogenesis, as NR5A1 (SF-1) is a master regulator for genes involved in cholesterol homeostasis.
• RUNX1 (Runt-related transcription factor 1): Sometimes appears in panels for comparison or in immune cell marker studies, though less directly related to cholesterol transport.

Other context-specific markers include:

• Cell type markers (CD45, F4/80 for macrophages, or DAPI for nuclei staining) to identify and distinguish specific populations in tissue or cellular experiments using 5A1.
• Other ABC transporter antibodies (such as ABCG1) for broader studies of the cholesterol efflux pathway.

These combinations are dictated by the focus of the experiment—cholesterol metabolism, cardiovascular biology, or extracellular matrix integrity studies most commonly use 5A1 (ABCA1) with ATP5A1, COL5A1, and NR5A1 antibodies in multiplex or comparative assays.

Clone 5A1 is a monoclonal antibody primarily cited in scientific literature for its ability to neutralize colony stimulating factor 1 (CSF-1, also known as M-CSF), an essential growth factor involved in the regulation of monocytes, macrophages, and related progenitor cells.

Key findings from scientific literature using clone 5A1 include:

• Neutralization of CSF-1 Activity: Clone 5A1 binds to mouse CSF-1 and inhibits its biological functions, including colony formation by tissue mononuclear phagocytes and the regulation of monocyte/macrophage lineage survival, proliferation, and differentiation.

• Macrophage-Specific Effects: Consistent findings show that 5A1 does not exert cytotoxic effects on breast carcinoma cells, but inhibits macrophage proliferation by inducing apoptosis in these cells. This selectivity is crucial for models investigating the role of macrophages in disease.

• Inhibition of Tumor Cell Invasion: The 5A1 antibody reduces macrophage-induced invasion of carcinoma cells in co-culture systems, such as in breast cancer models, supporting the role of CSF-1-mediated macrophage–tumor interactions in tumor progression. This effect was observed both with monocyte-derived macrophages and, to a lesser extent, with microglia.

• No Direct Effect on Cancer Cell Viability: Studies show clone 5A1 does not affect the viability, proliferation, or migration of breast cancer cells directly, isolating its effect to CSF-1-dependent processes, mainly in myeloid cell populations.

• Research Applications: Clone 5A1 is widely used in in vivo neutralization studies, mouse models of inflammation, cancer, and macrophage biology to dissect the functions of CSF-1 and its downstream cellular responses. Researchers commonly use this antibody to deplete or inhibit macrophages in disease models to clarify their pathogenic contributions.

• Mechanistic Insights: The use of 5A1 has helped elucidate mechanisms of macrophage-driven processes in various pathologies, including cancer invasion and possibly inflammatory conditions or atherosclerosis, by blocking CSF-1 signaling.

If referring to unrelated uses of “clone 5A1”, such as in plasmid studies in Borrelia burgdorferi or other organisms, those are distinct and unrelated to the anti-CSF-1 antibody applications described above.

Summary Table: Major Findings Linked to Clone 5A1 (Anti-CSF-1)| Finding | Description ||---------------------------------------|------------------------------------------------------------------------------|| Target | Mouse CSF-1 (M-CSF) || Functional Effect | Neutralizes CSF-1, inhibits phagocyte colony formation || Cell/Tissue Specificity | Affects monocytes, macrophages (not carcinoma cells) || Use in Research | In vivo and in vitro CSF-1 depletion/neutralization, macrophage depletion || Impact on Tumor Models | Reduces macrophage-induced tumor invasion; does not impact cancer cell growth || Mechanism | Induces apoptosis in macrophages; no effect on microglia proliferation |

These findings establish clone 5A1 as a research-standard antibody for functional CSF-1 inhibition in mouse models, particularly in studies of macrophage-driven disease mechanisms.

Clone 5A1 is a monoclonal antibody targeting mouse colony-stimulating factor 1 (CSF1), also known as macrophage colony-stimulating factor (M-CSF). The dosing regimens for clone 5A1 in different mouse models can vary significantly based on the specific application and experimental design.

1. Anti-Tumor Models

• Dose and Frequency: In anti-tumor models such as MGS1, clone 5A1 is used at a dose of 200 μg per mouse, administered intraperitoneally (i.p.) every 3-4 days. This regimen has shown potent anti-tumor effects.
• Higher Dose: In some studies, a higher dose of 0.5 mg per mouse is referenced, though specific administration schedules are not detailed for this dosage.

2. Immune-Mediated Models (e.g., EAE - Experimental Autoimmune Encephalomyelitis)

• Dose and Frequency: In models studying immune-related diseases, such as EAE, prophylactic treatments with anti-CSF1 (clone 5A1) at 200 μg per dose, administered i.p. every other day until disease onset, and then daily, have been used.

3. Pancreatic Tumor Models

• Dose and Frequency: In pancreatic tumor models, mice were treated with anti-CSF1R (clone 5A1) at a dose equivalent to 10 mg/kg, administered i.p. every 4 days starting on day 3 post-tumor implantation.

4. General Application

• Implications: The varied dosing schedules suggest that the optimal regimen can depend on the specific disease model, the stage of disease onset, and the desired effect on myeloid cells or other immune components.

Overall, the dosing regimen for clone 5A1 can vary significantly based on the specific research question, model system, and desired outcome. Factors such as the timing of administration relative to disease onset and the specific application (e.g., anti-tumor vs. immune-mediated diseases) play crucial roles in determining the optimal dosing strategy.