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

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

Product No.: M350

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Clone
MFL3
Target
CD178
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
FasL, Fas Ligand, FasL, Apo-1 Ligand, CD95 Ligand, TNFSF6
Isotype
IgG
Applications
B
,
FC
,
IF
,
in vivo

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Armenian Hamster
Recommended Dilution Buffer
Immunogen
B6 mouse FasL cDNA-transfected baby hamster kidney (B6 FasL/BHK) cells
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
<0.5 EU/mg as determined by the LAL method
Purity
≥98% monomer by analytical SEC
>95% by SDS Page
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Pathogen Testing
To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM<sup>TM</sup> antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Country of Origin
USA
Shipping
Next Day 2-8°C
Applications and Recommended Usage?
Quality Tested by Leinco
FC The suggested concentration for this CD178 antibody, clone MFL3, for staining cells in flow cytometry is ≤ 0.25 μg per 106 cells in a volume of 100 μl. Titration of the reagent is recommended for optimal performance for each application.
Additional Applications Reported In Literature ?
B
IF
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
Clone MFL3 recognizes an epitope on mouse FasL.
Background
FasL antibody, clone AFS98, recognizes Fas ligand (FasL), also known as CD178, Apo-1 ligand, and CD95 ligand. FasL is a 40 kDa type II integral membrane protein that belongs to the tumor necrosis factor (TNF) superfamily. FasL is expressed by activated T cells and natural killer (NK cells)1-3. Binding of FasL to its receptor Fas (CD95, APO-1) induces apoptotic cell death in Fas-expressing target cells, contributing to anti-viral immunity. FasL also contributes to peripheral tolerance and the downregulation of immune responses through activation-induced autocrine and paracrine T cell death4. FasL is also found in the anterior chamber of the eye and on Sertoli cells in the testis, and is implicated in immune-privilege at these sites5,6. FasL also contributes to CD8 proliferation and neutrophil recruitment7,8. Soluble FasL (26 kDa) can be released following cleavage by metalloproteinases and block FasL-mediated signaling9. Fas/FasL-signaling is involved in the development of many human diseases, including autoimmunity and cancer10. Many human tumors over-express FasL, resulting in tumor infiltrating lymphocyte (TIL) apoptosis and immune evasion, which is associated with poor prognosis11-14.
Antigen Distribution
FasL is expressed on activated T cells, NK cells, the eye, and testis.
Ligand/Receptor
Fas (CD95)
PubMed
NCBI Gene Bank ID
Research Area
Apoptosis
.
Cell Biology
.
Cell Death
.
Immunology
.
Neuroscience
.
Tumor Suppressors

Leinco Antibody Advisor

Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.

Clone MFL3 is most commonly used for in vivo blockade of Fas Ligand (FasL/CD178) in mice to inhibit FasL-mediated apoptotic signaling and modulate immune responses.

Key in vivo applications include:

  • Blocking FasL-mediated apoptosis: MFL3 inhibits the interaction between FasL (on activated T cells, NK cells, and some tissues) and its receptor Fas (CD95), thereby preventing cell death in contexts such as peripheral immune tolerance, tissue injury, and some disease models.
  • Modulation of immune responses: By interfering with FasL, MFL3 is used to study its roles in immune regulation, including T cell homeostasis, activation-induced cell death, and the resolution of immune responses.
  • Tumor immunology studies: Blocking FasL can be used to investigate mechanisms of tumor immune evasion, since some tumors overexpress FasL to induce apoptosis of infiltrating cytotoxic lymphocytes.
  • Studies of inflammation: MFL3 is used in models of autoimmunity, graft rejection, and inflammatory diseases to dissect the contribution of FasL-induced apoptosis in these processes.
  • Research on neutrophil function: FasL/Fas interactions are implicated in neutrophil extravasation, chemotaxis, and survival; thus, MFL3 allows in vivo experiments probing neutrophil biology.

Other technical applications in vivo:

  • Functional assays to test FasL-dependent cytotoxicity or survival.
  • Immunophenotyping via flow cytometry and immunohistochemistry in tissues to assess FasL expression/blockade effects.

Summary Table: Common In Vivo Applications of Clone MFL3 in Mice

ApplicationPurpose
FasL blockadePrevent apoptosis via Fas-FasL pathway
Study immune regulationAnalyze T cell and immune homeostasis
Tumor immunologyInvestigate tumor immune evasion via FasL
Inflammation modelsExamine FasL’s role in autoimmunity or tissue injury
Neutrophil biologyAssess FasL effects on neutrophil movement/survival

MFL3 is validated across mouse strains and is formulated for high purity, ultra-low endotoxin, and specific in vivo compatibility to minimize off-target or immunogenic effects in mouse models.

Based on research applications, MFL3 is commonly used in combination with several other antibodies and proteins to investigate immune cell function and apoptosis pathways.

T Cell Activation Studies

MFL3 is most frequently paired with anti-CD3 antibodies when investigating T cell activation. The anti-CD3 antibody clone 145-2C11 is a well-characterized reagent used to stimulate T cells, and researchers combine it with MFL3 to study the downstream effects of T cell activation on FasL expression and Fas-mediated apoptosis.

Apoptosis Pathway Research

For studying apoptotic mechanisms, MFL3 is commonly used alongside anti-Fas (CD95) antibodies. Since MFL3 targets the Fas Ligand (CD178), which binds to its receptor CD95/Fas to induce apoptotic cell death, pairing these antibodies allows researchers to comprehensively examine both sides of this critical death receptor-ligand interaction. This combination is particularly valuable for investigating how Fas/FasL interactions participate in T cell development, immune response regulation, and cell-mediated cytotoxic mechanisms.

Flow Cytometry Applications

In flow cytometry experiments, MFL3 is paired with a range of fluorescently labeled antibodies for detailed immune cell analysis. The antibody is available conjugated to various fluorophores including PE, APC-eFluor, and FITC, making it compatible with multi-color flow cytometry panels that include markers for different T cell subsets, activation markers, and other immune cell populations.

The key findings from scientific literature citing clone MFL3 focus on its specificity and functional use in mouse immunology research:

  • Specificity for Mouse Fas Ligand (FasL, CD178): Clone MFL3 is a monoclonal antibody that selectively reacts with mouse FasL, which is a member of the TNF superfamily expressed on activated T cells, natural killer (NK) cells, and in immune-privileged sites such as the eye and testis.
  • Functional Blocking and Detection: MFL3 is widely used to both detect and functionally block FasL in mice. This enables researchers to study the biological effects of FasL, particularly its role in Fas-mediated apoptosis through its interaction with the Fas (CD95) receptor.
  • Experimental Applications: The antibody is commonly employed in flow cytometry for the analysis of FasL-expressing cells and has also been used in in vivo and in vitro functional assays to elucidate the role of FasL in immune regulation, cell death, and disease models (e.g., autoimmunity, graft rejection, and immune privilege).
  • Blocking Studies: Literature reports that MFL3 can effectively block FasL function, as evidenced in experiments investigating immune mechanisms (e.g., reversal of T cell effects via FasL blockade).
  • Pathogen and Endotoxin Testing: Commercial preparations of MFL3 are stringently tested to ensure purity and absence of endotoxins and pathogens, supporting reliable experimental outcomes.

These findings make MFL3 an essential tool in studies exploring murine FasL biology, apoptosis, and immune modulation, with broad applications across immunology and related biomedical research.

Dosing Regimens of Clone MFL3 (Anti-Mouse Fas Ligand/CD178) Across Mouse Models

Clone MFL3 is a monoclonal antibody that targets mouse Fas ligand (FasL, CD178), a protein involved in apoptosis and immune regulation. While its biological effects—such as blocking apoptosis—have been demonstrated in specific experiments, the dosing regimens for MFL3 are not standardized across different mouse models and research applications.

General Considerations

  • Lack of Standardization: Published dosing schedules for MFL3 specifically are rare, and there are no widely accepted or universally applied protocols. This contrasts with other widely used in vivo monoclonal antibodies (e.g., anti-PD-1, anti-CTLA-4), where established dose ranges and administration schedules are commonly referenced.
  • Context-Dependent Use: Dosing is likely to be model-dependent, varying with the experimental design, disease context (e.g., tumor, autoimmune, or infection models), and the specific biological endpoint being studied (e.g., survival, immune cell infiltration, apoptosis inhibition).
  • Route of Administration: While not explicitly stated for MFL3, in vivo studies with monoclonal antibodies in mice typically use intraperitoneal (IP) injection as the standard route. However, this may vary based on the experimental needs.

Published Example

  • Cisplatin-Induced Lethality Model: One published study treated SCID/beige mice with MFL3 to block FasL and observed a significant restoration of survival after a lethal dose of cisplatin. However, this report does not specify the dose, frequency, or duration of MFL3 administration, highlighting a common gap in the literature for this clone.
  • Vendor Guidance: Major antibody vendors (e.g., Bio X Cell, Leinco Technologies) provide MFL3 for research use but do not provide general dosing recommendations, noting that regimens should be determined by the researcher and optimized for each experimental model. This is in contrast to some other antibodies, where vendors or guidelines suggest starting doses (e.g., 100–200 µg per mouse, every 3 days).

Best Practices for Determining MFL3 Dosing

  • Pilot Studies: Due to the lack of standardized regimens, researchers should conduct dose-finding studies to establish an effective and non-toxic dose for their specific mouse model and experimental conditions.
  • Emulate Established Protocols: For antibodies without established dosing, it is reasonable to start with dose ranges and schedules similar to those used for other monoclonal antibodies in mice (e.g., 100–200 µg per mouse every 3–4 days, IP), and adjust based on preliminary data.
  • Monitor Outcomes: Effectiveness (e.g., inhibition of apoptosis, changes in survival, immune cell profiles) and potential toxicity should be carefully monitored to refine the dosing regimen.

Summary

There is no uniform dosing regimen for clone MFL3 across different mouse models due to insufficient published data and vendor guidance. Researchers should design model-specific pilot experiments to determine optimal MFL3 doses and schedules, drawing on general principles for monoclonal antibody administration in mice when necessary. Always consult the primary literature and consider collaboration with laboratories that have previously used MFL3 in similar contexts for protocol refinement.

References & Citations

1. Okumura K., et al. (1994) Proc Natl Acad Sci USA. 91:4930–4934
2. Nagata S., et al. (1995) J Immunol. 154:3806–3813
3. Saito T., et al. (1995) J Exp Med. 181:1235–1238
4. Ferguson T A., et al. (1995) Science. 270:1189–1192
5. Duke R C., et al. (1995) Nature (London). 377:630–632
6. Fink PJ. (2000) J Immunol. 165(10):5537-43
7. Matsuzawa A., et al. (1998) J Immunol. 161: 4484–4488
8. Nagata S., et al. (1998) Nat Med. 4(1):31-6
9. Hueber AO., et al (2019) Cancers (Basel). 11(5):639
10. Kabelitz D., et al. (2000) Cancer Res. 60: 822–828
11. Giannarelli D., et al. (2000) Int J Cancer. 89: 127–132
12. Kanno H., et al. (2000) Br J Cancer. 82: 1446–1452.
13. Nagano H., et al. (Cancer) Br J Cancer. 82: 1211–1217
B
Flow Cytometry
IF
in vivo Protocol

Certificate of Analysis

Formats Available

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.