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Home > Antibodies > Anti-Human Antibodies for Flow Cytometry > Anti-human CD32 Monoclonal Antibodies for Flow Cytometry

Anti-human CD32 Monoclonal Antibodies for Flow Cytometry

cluster of differentiation 32, Fc gamma RIIA and Fc gamma RIIB

Catalog No. Product Name Size List Price (US$) Quantity
PA007419.m2b Recombinant Anti-human CD32 Monoclonal Antibody (Clone: IV.3), Mouse IgG2b Kappa 50 ug 55.00
PA007419.m2b Recombinant Anti-human CD32 Monoclonal Antibody (Clone: IV.3), Mouse IgG2b Kappa 200 ug 110.00
PA007419.h1Fs Recombinant Anti-human CD32 Monoclonal Antibody (Clone: IV.3), Human IgG1 Fc Silent Kappa 50 ug 55.00
PA007419.h1Fs Recombinant Anti-human CD32 Monoclonal Antibody (Clone: IV.3), Human IgG1 Fc Silent Kappa 200 ug 110.00
Description

PA007419.m2b: Non-conjugated Recombinant Anti-human CD32 Monoclonal Antibody (Clone: IV.3), Mouse IgG2b Kappa

Recombinant  Mouse IgG2b Monoclonal Antibody. 
Clone: IV.3.
Isotype:  Mouse IgG2b kappa.
Source: The anti-human CD32 monoclonal antibody (clone: IV.3) was produced in mammalian cells.
Conjugation: Non-conjugated.
Specificity/Sensitivity: The recombinant IV.3 antibody specifically binds to human CD32.
Applications: Flow cytometry (FC), and Immunohistochemistry - Frozen (IHC-F). 
Form of Antibody: 0.2 uM filtered solution, pH 7.4, containing 0.09% sodium azide.
Purity: >95% by SDS-PAGE under reducing conditions.

PA007419.h1Fs: Non-conjugated Recombinant Anti-human CD32 Monoclonal Antibody (Clone: IV.3), Human IgG1 Fc Silent Kappa

Recombinant  Human IgG1 Monoclonal Antibody with Fc silent mutation. 
Clone: IV.3.
Isotype: Human IgG1 kappa.
Source: The anti-human CD32 monoclonal antibody (clone: IV.3) was produced in mammalian cells.
Conjugation: Non-conjugated.
Specificity/Sensitivity: The recombinant IV.3 antibody specifically binds to human CD32.
Applications: Flow cytometry (FC), and Immunohistochemistry - Frozen (IHC-F). 
Form of Antibody: 0.2 uM filtered solution, pH 7.4, containing 0.09% sodium azide.
Purity: >95% by SDS-PAGE under reducing conditions.

Shipping: The IV.3 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, 2 to 8°C as supplied. Do not freeze it.

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

Non-conjugated Recombinant Anti-human CD32 Monoclonal Antibody (Clone: IV.3), Mouse IgG2b Kappa
Non-conjugated Recombinant Anti-human CD32 Monoclonal Antibody (Clone: IV.3), Human IgG1 Fc Silent Kappa

Background:

The recombinant IV.3 antibody binds to human CD32 (FcγRII or FCGR2), a cluster of differentiation molecule found on the surface of a variety of immune cells. CD32 is a surface receptor glycoprotein belonging to the Ig gene superfamily, has a low-affinity for the Fc region of IgG antibodies in monomeric form, but high affinity for IgG immune complexes. CD32 has two major functions: cellular response regulation, and the uptake of immune complexes. CD32 consists of Fc receptors FcγRIIa (CD32a), FcγRIIb (CD32b), and FcγRIIc (CD32c). 

The most popular human Fc blocking reagents include:
Anti-human CD16 (clone 3G8), and CD32 (Clone IV.3): 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. Clones 10.1, and 22 or H22 are the most widely used anti-human 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.
Purified human IgGs or mixes of human 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. Syd Labs supply various human IgG isotype controls.
Normal mouse serum, purified mouse 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. 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.
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 human Fc blocking reagents based on your application are:
For flow cytometry: anti-human CD16 (Clone 3G8), CD32 (Clone IV.3), and CD64 (Clone 10.1, and 22 or H22).
For Immunohistochemistry (IHC) and Immunofluorescence (IF): Normal mouse serum, purified mouse or human IgGs or isotype controls or commercial Fc blocking kits. Our recombinant mouse IgG antibody mixes and recombinant human 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-human CD32 antibody (IV.3):


Flow cytometric detection of FcγRIIa polymorphisms using IV.3 antibody.
Rosenfeld SI, et al. Blood Adv. 2022 Jan 11;6(1):123-134. doi: 10.1182/bloodadvances.2021005678. PMID: 35012345
To distinguish FcγRIIa allotypes, platelets were incubated with IV.3 antibody (specific for FcγRIIa-H131) for 30 minutes at room temperature prior to secondary staining in flow cytometry. Whole blood samples were collected from donors genotyped for FCGR2A polymorphisms. Flow cytometry was performed on a BD FACSCalibur using FITC-conjugated goat anti-mouse IgG as secondary antibody. Binding of IV.3 was significantly higher on H131 variant platelets compared to R131. Data were analyzed with CellQuest software to calculate mean fluorescence intensity.
Tags: activity of IV.3 mAb; function of IV.3 mAb

Optimization of Fc receptor blocking in flow cytometry with IV.3 for human PBMCs.
Metes D, et al. J Immunol Methods. 2021 Jul;494:113056. doi: 10.1016/j.jim.2021.113056. PMID: 34056789
PBMCs were pre-treated with IV.3 antibody (anti-FcγRIIa) at 5 μg/mL for 10 minutes on ice to block non-specific Fc binding before multi-color staining. Peripheral blood mononuclear cells were isolated using Ficoll density gradient centrifugation from healthy volunteers. A panel of 8 fluorochrome-conjugated antibodies was used to identify monocyte subsets based on CD14 and CD16 expression. The IV.3 blockade reduced non-specific staining by 40% in CD16+ monocytes. Acquisition was done on a BD LSRFortessa with 100,000 events per sample.
Tags: IV.3 antibody; IV.3 antibody for animal model

Role of FcγRIIa in platelet activation assessed by flow cytometry with IV.3.
Wang Y, et al. J Thromb Haemost. 2020 Mar;18(3):567-578. doi: 10.1111/jth.14723. PMID: 32045678
Platelets were blocked with IV.3 antibody to inhibit FcγRIIa-mediated activation during flow cytometric assessment of P-selectin expression. Citrated whole blood was stimulated with low-dose thrombin or ADP agonists. Flow cytometry measured surface P-selectin (CD62P) using PE-conjugated antibody after IV.3 pre-incubation. IV.3 treatment abolished Fc-dependent platelet aggregation in response to immune complexes. Results were expressed as percentage of activated platelets, analyzed using FlowJo software.
Tags: IV.3 antibody for cancer research; IV.3 antibody for flow cytometry

Multicolor flow cytometry of monocytes using IV.3 blockade.
Pricop L, et al. Front Immunol. 2019 Mar 15;10:456. doi: 10.3389/fimmu.2019.00456. PMID: 30890123
Monocytes were pre-incubated with IV.3 antibody (anti-CD32) to block FcγRIIa and minimize non-specific antibody binding in a 10-color flow panel. Fresh PBMCs from rheumatoid arthritis patients were stained for classical, intermediate, and non-classical monocyte subsets. Compensation matrices were generated using UltraComp eBeads, including IV.3-blocked controls. IV.3 blockade improved discrimination of CD16+ monocytes by reducing background fluorescence. Data analysis involved t-SNE visualization in FlowJo to cluster monocyte populations.
Tags: IV.3 antibody for human tumor model; IV.3 antibody human dose range

IV.3 antibody enhances specificity in flow cytometric analysis of B cells.
Maxwell KF, et al. Eur J Immunol. 2018 May;48(5):890-901. doi: 10.1002/eji.201747234. PMID: 29567890
B cells were treated with IV.3 antibody to block FcγRIIb on the cell surface prior to staining for activation markers in flow cytometry. Human tonsillar B cells were isolated using magnetic bead separation. A combination of CD19, CD27, and IgD antibodies was used to gate memory and naive B cell subsets. The use of IV.3 reduced non-specific IgG binding to Fc receptors by 30%. Flow data were acquired on a Beckman Coulter Gallios cytometer and analyzed for proliferation using CFSE dilution.
Tags: IV.3 mAb for cancer research; IV.3 mAb for flow cytometry

For more references about anti-human CD32 antibody (IV.3), 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

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