Anti-Mouse CD178 (FasL) – Purified in vivo PLATINUM™ Functional Grade

Clone MFL3 is a monoclonal antibody used in in vivo mouse studies primarily to target and block mouse Fas Ligand (FasL, CD178/CD95 Ligand), which is involved in inducing apoptotic cell death via interaction with the Fas receptor (CD95).

In vivo applications include:

• Blocking FasL-induced apoptosis: MFL3 can efficiently inhibit the cytotoxic activity mediated by FasL, thereby preventing apoptosis triggered through the CD178/CD95 pathway.
• Studying immune regulation: By inhibiting FasL, researchers use MFL3 in mouse models to investigate mechanisms of T cell development, regulation of immune responses, cell-mediated cytotoxicity, and autoimmune conditions driven by impaired Fas signaling.
• Tumor immunology studies: Some tumors over-express FasL to avoid immune detection; using MFL3 allows researchers to explore tumor immune evasion by blocking this pathway.
• Flow cytometry and cell staining: MFL3 is used to detect FasL expression on mouse cells, often following in vitro activation with anti-mouse CD3 antibodies, and can be utilized for cell sorting and phenotyping.

Typical Procedures:

• MFL3 is administered to mice (often as a low-endotoxin formulation) to block FasL function systemically or in specific tissues.
• For ex vivo staining (e.g., flow cytometry), MFL3 is used to detect and quantify FasL expression on activated T cells, sometimes amplified with secondary antibodies and bright fluorophores due to FasL's low density on cell surfaces.

Key points of use:

• The antibody specifically reacts with mouse FasL and does not cross-react with rat leukocytes.
• It is widely used for both functional blockade (in vivo/in vitro) and detection (flow cytometry/immunoassays).
• The reported applications include inhibition of FasL-mediated cell death, assessment of lymphocyte activation and immune regulation, and analysis of tumor-immune interactions.

This makes MFL3 a fundamental tool for in vivo dissection of FasL-dependent pathways in mouse models of immunity, autoimmunity, and cancer.

Sterile packaged monoclonal antibody clones like MFL3 should typically be stored at 2°C to 8°C (refrigerator temperature) for short-term storage (up to a few weeks) and at -20°C or lower for long-term storage to maintain stability and efficacy. For maximum preservation, long-term storage at -80°C is sometimes recommended, especially for periods exceeding one year.

• Short-term storage (up to a few weeks): 2°C to 8°C
• Long-term storage: -20°C or lower
• Very long-term storage: -80°C recommended for highest stability

Refrigeration at 2–8°C is standard for most monoclonal antibodies; avoid repeated freeze-thaw cycles, as these can degrade protein activity. If the sterile packaging is specifically designed for shelf-stable storage, and not just for contamination prevention, then packaging guidelines may override these standards, but in the absence of clone- or supplier-specific instructions, these temperature ranges should be followed.

If you require the absolute manufacturer guidance for MFL3, consult the technical data sheet included with your clone or the supplier’s official website, as antibody storage requirements can vary by formulation and packaging.

The MFL3 antibody targets mouse Fas Ligand (CD178, FasL, Apo-1 ligand, CD95 ligand), commonly used to study apoptosis, immune regulation, and T-cell biology. In published literature, MFL3 is routinely used alongside certain other antibodies and proteins, often to characterize cell subsets or analyze apoptotic pathways.

Commonly used antibodies/proteins in combination with MFL3 include:

• Anti-CD95 (Fas): Used to detect Fas receptor, often paired with MFL3 to study Fas/FasL interactions in apoptosis and immune regulation.

• Cell surface markers:

  • CD3: Marker for T cells, useful for gating activated T cells expressing FasL.
  • CD4/CD8: To identify T cell subsets that express or respond to FasL.
  • CD45: A pan-leukocyte marker for broader immune profiling.

• Apoptosis detection reagents:

  • Annexin V: Used to monitor early apoptosis, frequently co-stained with FasL (MFL3) and Fas.
  • Caspase activity probes/antibodies: Detection of caspase activation downstream of Fas signaling.

• Activation markers or stimulatory antibodies:

  • Anti-CD3 (e.g., clone 145-2C11): Used to activate T cells, which upregulate FasL and can be detected by MFL3.
  • Anti-CD28: Costimulatory molecule, applied in protocols that stimulate FasL expression.

• Cytokine detection antibodies:

  • TNF-alpha, IL-2, IFN-gamma: Common in immune studies where FasL and its role in T-cell-mediated cytotoxicity or regulation is investigated.

• Secondary antibodies conjugated to various fluorochromes: Used in indirect immunofluorescence to reveal MFL3 binding or co-localize additional markers.

These combinations allow researchers to study:

• Cell death pathways, by tracking both ligand (FasL/MFL3) and receptor (Fas/CD95).
• Immune cell activation and subset characterization, especially by co-staining T-cell markers and FasL.
• Cytokine profiles alongside FasL expression in immune response and autoimmune disease models.

Additionally, protocols often use isotype controls and live-dead cell exclusion dyes to ensure specificity in flow cytometry and immunofluorescence.

Alternative or additional antibodies depend on the experiment but most often include markers for immune cells (CD3, CD4, CD8), apoptosis-specific reagents (anti-Fas, annexin V), and activation/cytokine detection (e.g., anti-TNF-alpha, anti-IFN-gamma).

Key findings from scientific literature citing clone MFL3 center on its specificity for mouse Fas Ligand (CD178, FasL, Apo-1 ligand, CD95 ligand) and its role in studies of apoptosis and immune regulation.

• Blocking FasL-induced apoptosis: The MFL3 monoclonal antibody has been reported to block Fas Ligand (CD178)-induced apoptosis, making it a key reagent in experiments studying the function of FasL in promoting cell death, particularly in the context of T cell development, immune response regulation, and cell-mediated cytotoxic responses.

• Experimental applications: MFL3 is widely used in flow cytometry (typical staining concentration ? 0.25 ?g per 10^6 cells per 100 ?l) to detect mouse Fas Ligand, which is expressed by activated T cells, NK cells, and certain immune-privileged tissues (e.g., eye, testis). Its utility is in functional studies assessing FasL expression and function in mouse models.

• Contribution to peripheral tolerance: By blocking FasL, MFL3 enables investigations into FasL's role in peripheral tolerance and downregulation of immune responses, particularly through activation-induced T cell death (autocrine and paracrine mechanisms).

• Mechanistic studies: MFL3 is referenced in foundational studies mapping FasL’s influence on apoptotic signaling, thus contributing to our understanding of antiviral immunity, autoimmunity, and immune privilege.

No evidence was found in the search results linking MFL3 to research outside Fas Ligand studies (e.g., it is unrelated to the microglia clone "MGL3" discussed in neurodevelopmental clonal tracing). Thus, literature citing MFL3 focuses overwhelmingly on immunology, apoptosis, and mouse FasL functional analysis.