Catalog No.	PA007389.r2b
Product Name	Anti-mouse CD16/CD32 Antibody (Clone: 2.4G2) | PA007389.r2b
Supplier Name	Syd Labs, Inc.
Brand Name	Syd Labs
Synonyms	Mouse Fc Receptor Blocking Antibody, Mouse 2.4G2 Fc Block, Mouse CD16/CD32 Fc Block
Summary	The anti-mouse CD16/CD32 monoclonal antibody (clone: 2.4G2) was produced in mammalian cells.
Clone	2.4G2
Isotype	Rat IgG2b Kappa
Specificity/Sensitivity	The in vivo grade recombinant rat monoclonal antibody (clone: 2.4G2) specifically binds to mouse CD16 and CD32.
Applications	ELISA, flow cytometry, neutralization, functional assays such as bioanalytical PK and ADA assays, and those assays for studying biological pathways affected by the mouse CD16 and CD32 protein. Mouse Fc receptor blocking solution, mouse Fc receptor blocking reagent, and mouse Fc receptor blocking antibody.
Form Of Antibody	0.2 uM filtered solution, pH 7.4, no stabilizers or preservatives.
Endotoxin	< 1 EU per 1 mg of the protein by the LAL method.
Purity	>95% by SDS-PAGE under reducing conditions.
Shipping	The antibody is shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70°C as supplied. 1 month from date of receipt, 2 to 8°C as supplied.
Note	The mouse CD16/CD32 Fc blocking antibody (2.4G2 Fc blocking antibody) is an anti-mouse Fc receptor antibody for mouse FcR blocking reagent. Various isotypes are available for certain applications. Condition of sample preparation and optimal sample dilution should be determined experimentally by the investigator.
Order Offline	Phone: 1-617-401-8149 Fax: 1-617-606-5022 Email: message@sydlabs.com Or leave a message with a formal purchase order (PO) Or credit card.

Description

PA007389.r2b: Recombinant Anti-mouse CD16/CD32 Monoclonal Antibody (Clone: 2.4G2) , Rat IgG2b Kappa, In vivo Grade

The anti-mouse CD16/CD32 monoclonal antibody (clone: 2.4G2) was produced in mammalian cells.

Background

The recombinant 2.4G2 antibody binds to mouse CD16 (FcγIII) and CD32 (FcγII) receptor proteins. The major component of some commercial Fc block reagents is the 2.4G2 antibody. The rat, mouse, and human IgG Fc binds to mouse CD64 (FcγI). Thus, the recombinant 2.4G2 antibodies can be used to block non-specific binding of IgGs to mouse Fc receptors in flow cytometry, immunohistochemistry, immunoprecipitation, and blocking assays. Customers may choose different IgG isotypes for their own applications.

The most popular mouse Fc blocking reagents include:
Anti-Mouse CD16/CD32 (Clone 2.4G2): for flow cytometry, immunohistochemistry (IHC), and immunoprecipitation (IP). It is highly specific for FcγRII (CD32) and FcγRIII (CD16), and reduces background staining; it may not block all Fc receptors. Clone X54-5/7.1 is the most widely used anti-mouse FcγRI (CD64) clone for Fc blocking in flow cytometry and functional assays. It prevents non-specific Fc receptor binding on monocytes, macrophages, and dendritic cells, improving antibody specificity.
Normal mouse serum, purified mouse IgGs from mouse serum, or mixes of mouse isotype controls: for general Fc blocking in IHC, and immunofluorescence (IF). It is cheap and easy to use but it is less specific, and may even introduce unwanted immunoglobulins. Our recombinant mouse IgG antibody mixes are affordable; the ratio of various IgGs is optimized and adjustable without any unwanted immunoglobulins.
Purified rat IgG or isotype controls: for flow cytometry. It can serve as a control to block Fc receptors but it is non-specific for Fc receptors. Syd Labs supply various rat IgG isotype controls.
Bovine Serum Albumin (BSA) or FBS: for general blocking in Western Blot (WB) and IHC. They are readily available to reduce non-specific binding but less effective at Fc receptor blocking.
Commercial Fc blocking solutions: for flow cytometry, IHC, and functional assays. They are normally pre-optimized and highly effective but more expensive than DIY solutions.

Best choice of mouse Fc blocking reagents based on your application are:
For flow cytometry: anti-mouse CD16/CD32 (Clone 2.4G2).
For Immunohistochemistry (IHC) and Immunofluorescence (IF): Normal mouse serum, purified mouse or rat IgGs or isotype controls or commercial Fc blocking kits. Our recombinant mouse IgG antibody mixes and recombinant rat IgG isotype controls are affordable; the ratio of various IgGs is optimized and adjustable without any unwanted immunoglobulins.
For Western Blot and ELISA: BSA or FBS.
For functional assays (e.g., blocking Fc-mediated effects): Commercial Fc blocking reagents.

References of Anti-mouse CD16 / CD32 Monoclonal Antibodies:

1、Anti-FcγRIIB mAb suppresses murine IgG-dependent anaphylaxis by Fc domain targeting of FcγRIII.
Clay, C. D., et al. J Allergy Clin Immunol. 2018 Apr;141(4):1373-1381.e5. doi: 10.1016/j.jaci.2017.05.027. PMID: 28624610.
“We previously characterized a mouse model of IgG-mediated anaphylaxis triggered by 2.4G2, a rat IgG2b mAb that directly binds to and blocks both FcγRIII and FcγRIIB but also inhibits stimulation of mouse myeloid cells through FcγRI and FcγRIV. …Our data suggest that 2.4G2 inhibition of FcγRI and FcγRIV results from an interaction between 2.4G2’s Fc domains and these receptors. …K9.361 and 2.4G2 mAbs were digested for 7.5 hours at 37°C with immobilized pepsin (Thermo Scientific) while being rotated and then purified with the NAb Protein A Plus column (Thermo Scientific) and concentrated with centrifugal filter devices (Amicon, EMD Millipore, Norwood, Ohio). …Our previous studies demonstrated that 2.4G2 (an mAb that binds to both the stimulatory FcγRIII and the inhibitory FcγRIIB) induces FcγRIII-dependent anaphylaxis that is manifested as shock-associated hypothermia. …Taken together, these observations suggest that induction of anaphylaxis by anti-FcγRIIB mAb requires 2 interactions: with FcγRIIB through its antigen-binding site and with FcγRIII through its Fc domain.”

2、Enhanced uptake of blood coagulation factor VIII containing immune complexes by antigen presenting cells.
Hartholt, R. B., et al. J Thromb Haemost. 2017 Feb;15(2):329-340. doi: 10.1111/jth.13570. PMID: 27868337.
“BMDCs were able to efficiently internalize FVIII‐IC in a dose‐dependent manner, 3–4‐fold more efficiently when compared with equimolar concentrations of non‐complexed FVIII. Uptake of FVIII‐IC, but not FVIII alone, could be inhibited with anti‐Fcγ receptor (FcγR) antibody 2.4G2, indicating functional involvement of FcγR. …IgG and CD206 were stained in TBS supplemented with 0.5% BSA, 1 : 500 mouse Fc block (2.4G2, Miltenyi Biotech) and 0.05% saponine. …Therefore, we incubated BMDCs with 10 μg mL−1 monoclonal antibody 2.4G2, which interferes with the binding of IgG to FcγRIIb and FcγRIII. …The uptake of FVIII was not reduced in the presence of 2.4G2 (Fig. 3C). These results indicate that Fcγ receptors mediate the uptake of FVIII‐IC by BMDCs. …The development of FVIII inhibitors precludes further treatment with FVIII; bleeding episodes in these patients are managed by FVIII bypassing therapies.”

3、Pre-Existing Humoral Immunity Enhances Epicutaneously-Administered Allergen Capture by Skin DC and Their Migration to Local Lymph Nodes.
Hervé, P.-L., et al. Front Immunol. 2021 Mar 26;12:609029. doi: 10.3389/fimmu.2021.609029. PMID: 33868229.
“In line with these observations, our recent preliminary data showed that injection of 2.4G2 blocking antibody prior to patch application had no impact on the subsequent modulation of PD-L2 and CD86 expression in OVA-positive DCs. …In these future experiments, it would also be relevant to include additional groups of control mice that receive “blank” epicutaneous patches (containing excipient without OVA) to avoid any potential bias linked to patch application and/or excipient (PBS). …Our results strongly suggest that pre-existing antibodies did not interfere with the uptake of an epicutaneously-delivered antigen by skin DC, suggesting that the epicutaneous route, similar to the mucosal route of immunization, may alleviate the loss of vaccine efficacy in seropositive individuals. …If confirmed, this could give a solid advantage to epicutaneous delivery for boosting vaccine responses, and also suggest a role for this delivery route in priming of vaccine responses in seropositive individuals, such as young infants who have maternal antibodies. …To investigate the involvement of FcγR, animals received 2.4G2 (anti-FcγRII/RIII) blocking antibody, 24 hours before patch application.”

4、Suppression of Pathological Allergen-Specific B Cells by Protein-Engineered Molecules in a Mouse Model of Chronic House Dust Mite Allergy.
Ralchev, N., et al. Int J Mol Sci. 2024 Dec 20;25(24):13661. doi: 10.3390/ijms252413661. PMID: 39769423.
“We hypothesize that administration of a chimeric molecule, composed of a monoclonal 2.4G2 antibody against FcγRIIb coupled with a Der p1-derived peptide (Dp52–71), can selectively eliminate disease-associated allergen-specific B cells in a chronic mouse model of HDM allergy. …Dp52–71 and irrelevant chimeras were constructed by coupling rat 2.4G2 monoclonal antibodies with peptide epitopes from Der p1 or irrelevant peptides, respectively. …Both the Dp52–71and irrelevant chimeras bound to the surface of CD19 cells with the same intensity as the pure 2.4G2 antibody in PBS- and HDM-challenged mice. …We further analyzed the capacity of the chimeric molecules to compete against a commercial FITC-conjugated 2.4G2 antibody for binding to the FcγRIIb receptor. …Chimeric molecules consisting of a monoclonal 2.4G2 antibody that recognizes FcγRIIb coupled with proven epitope-carrying peptides (Dp52–71), and control chimeras with the same antibody, were constructed.”

For more references about Anti-mouse CD16 / CD32 Monoclonal Antibodies please contact our scientific support team with message@sydlabs.com.

Syd Labs provides the following in vivo grade recombinant anti-human CD16, CD32, and CD64 monoclonal antibodies:

Recombinant Anti-human CD16 monoclonal antibody (Clone: 3G8)
Recombinant Anti-human CD32 monoclonal antibody (Clone: IV.3)
Recombinant Anti-human CD64 monoclonal antibody (Clone: H22)

Syd Labs provides the following in vivo grade recombinant anti-mouse CD16, CD32, and CD64 monoclonal antibodies:

Recombinant Anti-mouse CD16/CD32 monoclonal antibody (Clone: 2.4G2)

Syd Labs provides the following recombinant anti-human CD16, CD32, and CD64 monoclonal antibodies for flow cytometry:

Recombinant Anti-human CD16 monoclonal antibody (Clone: 3G8) for flow cytometry
Recombinant Anti-human CD32 monoclonal antibody (Clone: IV.3) for flow cytometry
Recombinant Anti-human CD64 monoclonal antibody (Clone: H22) for flow cytometry

Anti-mouse CD16/CD32 Antibody ( 2.4G2) from: Recombinant Anti-mouse CD16/CD32 Monoclonal Antibody, Rat IgG2b Kappa (Clone: 2.4G2): PA007389.r2b Syd Labs

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