Anti-Mouse CD32/CD16 [Clone 2.4G2] — Purified in vivo GOLD™ Functional Grade
Anti-Mouse CD32/CD16 [Clone 2.4G2] — Purified in vivo GOLD™ Functional Grade
Product No.: C381
Clone 2.4G2 Target CD32/CD16 Formats AvailableView All Product Type Monoclonal Antibody Alternate Names Fcγ R III/II, Ly-17 Isotype Rat IgG2b Applications B , FA , FC , IHC FF , in vivo , IP , PhenoCycler® , WB |
Data
Antibody DetailsProduct DetailsReactive Species Mouse Host Species Rat Recommended Isotype Controls Recommended Isotype Controls Recommended Dilution Buffer Immunogen Sorted pre-B cells Product Concentration ≥ 5.0 mg/ml Endotoxin Level < 1.0 EU/mg as determined by the LAL method Purity ≥95% 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. 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 RRIDAB_2737484 Applications and Recommended Usage? Quality Tested by Leinco FC The suggested concentration for this 2.4G2 antibody for staining cells in flow cytometry is ≤ 1 μg per 106 cells in a volume of 100 μl. Titration of the reagent is recommended for optimal performance for each application. WB The suggested concentration for this 2.4G2 antibody for use in western blotting is 1-10 μg/ml. Additional Applications Reported In Literature ? CODEX® Additional Reported Applications For Relevant Conjugates ? B IP For specific conjugates of this clone, review literature for suggested application details. Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionDescriptionSpecificity Clone 2.4G2 recognizes the FcγIII and FcγII receptors. Background CD16 is expressed in two forms: CD16a and CD16b. CD16a (FcγRIIIA) is a 50-65 kD polypeptide-anchored transmembrane protein. CD16b (FcγRIIIB) is a 48 kD GPI-anchored protein whose extracellular domain is over 95% homologous to that of CD16a. CD16 regulates both phagocytosis and antibody-dependent cell-mediated cytotoxicity. It has been reported that CD16 is involved in Natural Killer Cell activation and plays a role in signal transduction. The receptors, CD32 (FcγRIII) and CD16 (FcγRII), are 40-60 kD and bind antibody-antigen immune complexes and mediate adaptive immune responses. Antigen Distribution These receptors are present on B cells, monocyte/macrophages, NK cells, neutrophils, mast cells and dendritic cells. Ligand/Receptor IgG Function Low affinity receptors for IgG PubMed UniProt.org Research Area Immunology . Innate Immunity Leinco Antibody AdvisorPowered 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 2.4G2 is utilized in in vivo mouse studies primarily as an Fc receptor blocking antibody to prevent non-specific binding and interference from endogenous antibodies during experimental procedures. Primary Mechanism and PurposeThe 2.4G2 antibody works by blocking the interaction between IgG antibodies and CD16 (Fc?RIII) or CD32 (Fc?RII) receptors on immune cells. These receptors are expressed on various immune cell types including B cells, monocytes, macrophages, NK cells, neutrophils, granulocytes, mast cells, and dendritic cells. By occupying these Fc receptor binding sites, the 2.4G2 antibody prevents non-specific binding of experimental antibodies that could interfere with study results. Specialized In Vivo FormulationsFor in vivo applications, specialized formulations of the 2.4G2 antibody have been developed with specific characteristics that make them suitable for use in living mice: Low Endotoxin Content: The in vivo grade formulations contain endotoxin levels of ?0.01 EU/?g or <1 EU per 1 mg, as determined by the Limulus amebocyte lysate (LAL) test. This low endotoxin level is crucial to prevent inflammatory responses that could confound experimental results. Azide-Free Formulation: Unlike some laboratory antibodies that contain sodium azide as a preservative, the in vivo grade 2.4G2 is formulated without azide, making it safe for injection into living animals. Modified Variants for In Vivo UseSeveral engineered versions of the 2.4G2 antibody have been developed specifically for enhanced in vivo performance: LALA-PG Mutation: Some versions contain LALA-PG mutations in the Fc fragment, which render the antibody unable to bind to endogenous Fc? receptors. This modification prevents the blocking antibody itself from causing unwanted immune activation while still allowing it to block the target receptors. Species Switching: Chimeric versions have been created where the variable domains remain identical to the original 2.4G2, but the constant regions are switched from rat IgG2b to mouse IgG2a. This modification can reduce immunogenicity in mouse models. Applications in Mouse StudiesIn vivo applications of 2.4G2 include blocking Fc receptors during studies investigating antibody-mediated immune responses, preventing interference in flow cytometry analysis of tissues harvested from treated mice, and neutralizing endogenous Fc receptor activity in functional assays studying biological pathways affected by CD16 and CD32 proteins. The antibody is also used in bioanalytical pharmacokinetic (PK) and anti-drug antibody (ADA) assays in mouse models. The careful formulation and engineering of these in vivo grade 2.4G2 antibodies ensures they can be safely administered to mice while effectively blocking Fc receptors without causing adverse reactions or confounding experimental results. The correct storage temperature for sterile packaged clone 2.4G2 (anti-mouse CD16/CD32 monoclonal antibody) is 2–8°C for short-term (up to 1 month) and -20°C to -70°C for long-term storage (up to 12 months). It is important to avoid repeated freeze-thaw cycles and use a manual defrost freezer for long-term storage. Essential context:
Additional information:
Commonly Used Antibodies and Proteins with 2.4G2 in the LiteratureThe 2.4G2 monoclonal antibody is best known for its role in blocking mouse CD16 and CD32 (Fc?RIII and Fc?RII), preventing non-specific binding of the Fc portion of immunoglobulins during immunoassays such as flow cytometry and immunofluorescence. However, when used in research, 2.4G2 is not typically the primary immune marker; rather, it is employed as a blocking reagent to improve the specificity of other antibodies targeting cell surface markers. Below are the main categories of antibodies and proteins commonly used in concert with 2.4G2 in the literature. Common Antibody Partners in Multiparameter AnalysisIn mouse immunology and cell phenotyping studies, 2.4G2 is used alongside a variety of fluorescently labeled antibodies that target specific leukocyte subsets or activation markers. Examples from the literature include:
These antibodies are routinely used in panels for comprehensive immune cell profiling, where 2.4G2 is added as a blocking reagent to minimize non-specific Fc receptor-mediated staining before applying these primary antibodies. Blocking with 2.4G2 in Functional Assays
Other Proteins and Controls
Summary Table of Common Antibodies Used with 2.4G2
ConclusionIn the literature, 2.4G2 is most frequently used as a blocking reagent in combination with a wide array of cell surface marker antibodies (e.g., B220, CD3, CD19, CD11b, Gr1, NK1.1) for multiparameter flow cytometry and immunofluorescence in mouse samples. Appropriate isotype controls and careful selection of secondary antibodies (avoiding anti-rat IgG2b) are also standard practice. These combinations enable precise identification and analysis of immune cell subsets by minimizing non-specific staining through Fc receptor blockade. Clone 2.4G2 is a monoclonal antibody widely used in immunology research, especially in mouse models, and is most frequently cited for its ability to block or detect Fc gamma receptor II (CD32, Fc?RII) and Fc gamma receptor III (CD16, Fc?RIII). Key findings and scientific uses from the literature include:
In summary, the main scientific contribution of clone 2.4G2 is its reliable and efficient blocking of Fc?RII and Fc?RIII, which underpins much of mouse immunology research by allowing for more specific analysis of immune cell markers and functions. References & Citations1.) Titas, J. A. et al. (1982) J. Immunol. 133:556 2.) Rodewald, H. et al. (1992) Cell 69:139 3.) Skyberg, J. A. et al. (2020) Infection and Immunity. 88: 5 4.) Forte et al. (2020) Cell Reports. 30:3149–3163 Journal Link 5.) Forte, E. et al. (2020) Cell Reports 30(9):3149-3163.e6 Journal Link Technical ProtocolsCertificate of Analysis |
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