Anti-Mouse CD11a (Clone FD441.8) – Purified in vivo PLATINUM™ Functional Grade

Anti-Mouse CD11a (Clone FD441.8) – Purified in vivo PLATINUM™ Functional Grade

Product No.: C603

[product_table name="All Top" skus="C303"]

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Clone
FD441.8
Target
CD11a
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
LFA-1α
Isotype
Rat IgG2b
Applications
FA
,
FC
,
N
,
WB

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Isotype Controls
Recommended Dilution Buffer
Immunogen
Cloned CTL B18 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™ 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 FD441.8 antibody for staining cells in flow cytometry is ≤ .25 μg per 106 cells in a 100 µl volume or 100 µl of whole blood. Titration of the reagent is recommended for optimal performance for each application.
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 FD441.8 recognizes an epitope on mouse CD11a.
Background
LFA-1α (CD11a) and CD18 are the Integrin alpha-L and beta-2 chains respectively that combine to form LFA-1, a glycoprotein and a member of the Integrin family. Integrin alpha-L/beta-2 is a receptor for ICAM1, ICAM2, ICAM3, ICAM4 and for F11R. LFA-1 participates in the immunological synapses between CD8+ T lymphocytes and antigen-presenting cells. The absence of LFA-1α or ß may induce LAD. The antigen contributes to natural killer cell cytotoxicity, and is involved in various immune phenomena such as leukocyte-endothelial cell interaction, cytotoxic T-cell mediated killing, and antibody dependent killing by granulocytes and monocytes. The CD11b/CD18 antigen is a heterodimeric surface glycoprotein on leukocytes and belongs to the ß2 integrin family. CD11b functions as a receptor for C3bi complement, clotting factor X, fibrinogen and ICAM-1. CD11c forms an α/ß heterodimeric glycoprotein (CD11c/CD18 complex) which belongs to the ß2 integrin family. The complex binds fibrinogen and reportedly serves as a receptor for iC3b and ICAM-1. During inflammatory responses, it mediates cell to cell interaction and is important in both monocyte adhesion and chemotaxis.
Antigen Distribution
CD11a is present on thymocytes, blood lymphocytes, bone marrow cells and certain lymphoma and macrophage-like cell lines.
Ligand/Receptor
ICAM-1 (CD54), ICAM-2 (CD102), and ICAM-3 (CD50)
Function
LFA-1 plays a central role in leukocyte intercellular adhesion.
PubMed
NCBI Gene Bank ID
Research Area
Cell Adhesion
.
Cell Biology
.
Costimulatory Molecules
.
Immunology
.
Signal Transduction

Leinco Antibody Advisor

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Clone FD441.8 is a rat monoclonal antibody that targets mouse CD11a (also known as LFA-1α). It is commonly used in in vivo applications in mice for the following purposes:

  1. LFA-1 Neutralization: Clone FD441.8 is used to block or inhibit the function of LFA-1, which plays a crucial role in intercellular adhesion, leukocyte-endothelial cell interaction, and cytotoxic T-cell mediated killing. This is particularly useful in studies involving immune response and cell interactions.

  2. Flow Cytometry: Although not the primary application in vivo, it is suitable for flow cytometry (FC) when used in conjunction with other in vivo assays.

  3. Functional Assays: The antibody is used in functional assays to inhibit or block LFA-1 activity, which helps in studying its role in immune phenomena such as leukocyte adhesion and migration.

  4. Immunological Studies: By blocking LFA-1α, researchers can study its impact on various immunological processes, including T-cell interactions and immune cell trafficking.

In summary, Clone FD441.8 is primarily used for LFA-1 neutralization in in vivo studies, facilitating the investigation of immune functions and cell adhesion processes in mice.

Commonly Used Antibodies and Proteins with FD441.8 in the Literature

Anti-CD11a (FD441.8) is a rat monoclonal antibody specific for mouse CD11a (LFA-1α, also known as integrin alpha-L), a key component of the LFA-1 integrin complex involved in leukocyte adhesion and migration. In published research, FD441.8 is frequently used alongside other antibodies and proteins to study immune cell interactions, adhesion, and trafficking. Here are some of the most commonly co-used reagents and their specific roles:

Blocking, Functional, and Isotype Controls

  • Purified Rat IgG2b, κ Isotype Control (LTF-2): Often used as a negative control to ensure the specificity of FD441.8 in flow cytometry and functional assays.
  • Functional Blocking: FD441.8 itself is used for blocking LFA-1 interactions, for example, to study the role of LFA-1 in leukocyte adhesion or migration.
  • Preclinical Functional Grade Antibodies: These are designed for in vitro and in vivo studies, ensuring minimal endotoxin and protein contamination for immune modulation experiments.

Co-Used Monoclonal Antibodies for Costimulation Blockade

In the context of immune response modulation, especially in transplant and autoimmune research, FD441.8 (anti-mouse CD11a) is often paired with antibodies targeting costimulatory molecules to achieve synergistic immunosuppression:

  • Monoclonal Antibodies Against CD154 (e.g., MR-1): In studies of transplant tolerance and islet xenograft survival, FD441.8 has been used together with anti-CD154 (MR-1) to provide dual blockade of LFA-1 and CD40-CD154 pathways, significantly extending graft survival in mouse models. This combination targets both the antigen-independent and antigen-dependent pathways of T cell activation.
  • Other Integrin Blockers: While FD441.8 targets CD11a (α chain of LFA-1), complementary antibodies against the CD18 (β chain) are sometimes used to fully block LFA-1 function, though these are less frequently cited together with FD441.8 in the provided literature.

Antibody Panels for Flow Cytometry and Immunophenotyping

FD441.8 is often included in multi-color flow cytometry panels to identify and characterize immune cell subsets. While the specific combinations vary by experiment, common co-stains might include:

  • Anti-CD3, Anti-CD4, Anti-CD8: For T cell subset identification.
  • Anti-B220, Anti-CD19: For B cell identification.
  • Anti-CD44, Anti-CD62L: For memory/naïve T cell discrimination.
  • Anti-Mac-1 (CD11b): For myeloid lineage cells, sometimes analyzed alongside CD11a to distinguish leukocyte subsets.
  • Anti-α4β7 (DATK32), Anti-MAdCAM-1 (MECA-367): For studying gut-tropic lymphocytes and mucosal homing.

Functional Assays and Cell Migration Studies

  • Blocking Adhesion Molecules: In studies of leukocyte trafficking, FD441.8 is used to block LFA-1, while antibodies against P-selectin, MAdCAM-1, or other adhesion molecules may be used to dissect the contributions of different pathways.
  • In Vivo Models: For in vivo functional studies, FD441.8 is sometimes administered systemically, alone or in combination with other blocking antibodies, to assess the role of LFA-1 in disease models.

Summary Table: Key Antibodies and Proteins Used with FD441.8

Antibody/ProteinTargetCommon Use Case with FD441.8Reference
Anti-CD154 (MR-1)CD40 ligandCostimulation blockade, transplant tolerance
Rat IgG2b (LTF-2)Isotype controlFlow cytometry control
Anti-CD3, CD4, CD8T cell markersImmunophenotyping
Anti-B220, CD19B cell markersImmunophenotyping
Anti-CD44, CD62LMemory/naïve TImmunophenotyping
Anti-Mac-1 (M1/70)CD11bMyeloid cell identification
Anti-MAdCAM-1 (MECA-367)MAdCAM-1Mucosal homing studies
Anti-α4β7 (DATK32)α4β7 integrinGut-homing lymphocyte studies

Typical Experimental Contexts

  • Transplant Models: FD441.8 + anti-CD154 for costimulation blockade.
  • Flow Cytometry: FD441.8 in multi-color panels with lineage and activation markers.
  • Adhesion/Migration: FD441.8 combined with blockers of other adhesion molecules (e.g., P-selectin, MAdCAM-1) to dissect leukocyte trafficking mechanisms.
  • In Vivo Functional Studies: FD441.8 alone or with other functional grade antibodies for immune modulation.

Conclusion

FD441.8 is most commonly used with antibodies targeting costimulatory molecules (especially CD154), isotype controls, and a range of cell lineage markers for comprehensive immunophenotyping. Its use in blocking studies often involves complementary adhesion molecule blockers to dissect complex leukocyte trafficking pathways. The specific combination depends on the experimental question, but these pairings are well established in the immunology literature.

Key Scientific Findings Involving Clone FD441.8

FD441.8 is a rat monoclonal antibody (rat IgG2b isotype) specifically recognizing mouse CD11a (LFA-1α), a subunit of the LFA-1 (Lymphocyte Function-Associated Antigen-1) integrin that mediates leukocyte adhesion and immune synapse formation. It is widely used in immunological research for blocking LFA-1 function, flow cytometry, and related applications.

Below are the key findings and applications from the scientific literature citing clone FD441.8:

Functional Studies

  • LFA-1 Blockade: FD441.8 is well-characterized as a blocking antibody that inhibits LFA-1-mediated cellular adhesion. Its application has been shown to block ConA-facilitated lysis but does not affect PHA-facilitated lysis, highlighting specific context-dependent effects of LFA-1 blockade.
  • Immune Synapse Inhibition: FD441.8 has been used to demonstrate that LFA-1 is critical for the formation of immunological synapses between CD8+ T lymphocytes and antigen-presenting cells, a fundamental process in adaptive immunity.
  • Blockade of Antigen Presentation: In co-culture studies, FD441.8 specifically blocked antigen presentation by cells expressing ICAM-1 (the ligand for LFA-1), but not by cells lacking ICAM-1 expression, underlining the importance of LFA-1/ICAM-1 interaction for efficient T cell activation.
  • Naive B Cell Trafficking: FD441.8 has been employed to show that local chemokine availability directs naive B cell trafficking, and blockade of LFA-1 with this antibody modulates such trafficking, indicating a crucial role for LFA-1 in lymph node homing and immune surveillance.
  • Graft Infiltration Inhibition: In transplantation models, administration of FD441.8 was shown to inhibit early infiltration of endogenous leukocytes into allografts, suggesting therapeutic potential for LFA-1 blockade in reducing graft rejection.

Technical and Preclinical Applications

  • Antibody Specifications: FD441.8 is produced with very low endotoxin levels—making it suitable for in vivo functional studies—and is available as “purified” as well as “functional grade” for a range of applications including immunohistochemistry, immunoprecipitation, and more.
  • In Vivo Use: The antibody is validated for in vivo administration, where it can effectively inhibit LFA-1-dependent immune processes in mouse models.
  • Pharmaceutical-Grade Production: Manufacturers like Leinco Technologies produce animal-free, low-endotoxin versions of FD441.8, expanding its utility in preclinical research.

Summary Table: Major Biological Effects of FD441.8-Mediated LFA-1 Blockade

EffectExperimental ContextCitation
Blocks ConA-facilitated lysisIn vitro cytotoxicity assays
Inhibits immune synapse formationCD8+ T cell—APC interaction
Blocks antigen presentation via ICAM-1T cell activation assays
Modulates naive B cell traffickingLymph node homing models
Reduces leukocyte infiltration in graftsTransplantation models

Conclusion

Clone FD441.8 is a pivotal tool for dissecting LFA-1 (CD11a) function in murine immunology, with a strong track record in blocking leukocyte adhesion, immune synapse formation, and modulating lymphocyte trafficking. Its specific, functional blockade has provided direct evidence for the critical role of LFA-1 in immune responses, antigen presentation, and transplant rejection, and it remains widely used in both basic and translational research.

Dosing Regimens of clone FD441.8 (Anti-LFA-1) in Different Mouse Models

Clone FD441.8 is a monoclonal antibody targeting LFA-1 (CD11a/CD18), commonly used in mouse models to study immune responses, particularly in transplantation, inflammation, and immune cell trafficking research.

Experimental Transplant Model

In a cardiac allograft model, mice (C57BL/6 and A/J strains) received intraperitoneal injections of anti-LFA-1 mAb (clone FD441.8) at 0.2 mg (200 μg) per mouse on days –1 and 0, or days +3 and +4 post-transplant. This schedule was designed to transiently block LFA-1 during critical early phases of alloantigen recognition and graft rejection. Notably, this dosing was used alongside other immune-modulating antibodies and was shown to impact alloreactive T-cell responses and graft outcome.

Xenotransplantation/Pancreatic Tissue Model

In a pig-to-mouse pancreatic tissue transplantation study, dosing of anti-LFA-1 (clone FD441.8, from the same source and presumably the same preparation) was given intraperitoneally (i.p.) at 200 μg/mouse on days 0, 2, 4, and 6 post-transplant, with a single injection repeated biweekly until 3 months post-transplant. This regimen was part of a broader immune suppression protocol and aimed for sustained LFA-1 blockade to promote engraftment and prevent graft rejection.

Comparison Table

Model TypeDose per MouseSchedule (Post-Transplant)Key Findings/Context
Cardiac Allograft200 μg i.p.Days –1 & 0 or Days +3 & +4Blocks early alloreactive T-cell responses
Xenotransplantation200 μg i.p.Days 0, 2, 4, 6 & biweekly × 3 moSustained blockade for long-term engraftment

Key Observations

  • Total dose per injection remains consistent at 200 μg/mouse across both studies, but the dosing schedule varies depending on the experimental endpoints and model requirements.
  • Administration route is universally intraperitoneal (i.p.), which is standard for in vivo antibody delivery in mouse models.
  • Timing of administration is tailored to the biological question: acute blockade for allograft models (just before and after transplant), versus prolonged/repeated dosing for xenograft studies to maintain immunosuppression.
  • Combinatorial therapy is common; anti-LFA-1 is rarely used alone and is typically part of multi-agent immunosuppressive regimens.

Practical Considerations

  • Antibody preparation and source are critical; both cited studies used clone FD441.8 from Bio Express (now Bio X Cell), ensuring reagent consistency.
  • Mouse strain and age do not appear to affect the primary dosing regimen, as both C57BL/6 (commonly used in transplantation) and other strains were treated with the same dose.
  • No reports were found for significantly higher or lower doses in other published mouse models, suggesting 200 μg/mouse i.p. is a robust, empirically validated reference dose for FD441.8 in mice.

Summary

The dosing regimen for clone FD441.8 (anti-LFA-1) in mouse models is 200 μg per mouse, intraperitoneally, but the frequency and duration of administration vary according to the experimental model and desired outcome—acute dosing for short-term immune blockade or repeated dosing for sustained suppression. This flexibility allows researchers to tailor the regimen to the specific biological processes under investigation, while the per-dose amount remains consistent across published protocols.

References & Citations

1. Suratt, Benjamin T. et al. (2009) J Immunol.182(1): 604–612. PubMed
2. Wuthrich, R. P. et al. (1991) Cell Immunol. 144:22
3. Sanchez-Madrid, F. et al. (1983) J. Exp. Med. 158(20):586
4. Pont, S. et al. (1986) J. Immunol. 136(10):3750
FA
Flow Cytometry
N
General Western Blot Protocol

Certificate of Analysis

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