Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)

Rabbit Recombinant Monoclonal FCGR3 antibody. Suitable for ICC/IF, IP, Flow Cyt, WB, IHC-P and reacts with Mouse, Recombinant fragment - Mouse samples. Cited in 4 publications.

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Images

Flow Cytometry - Anti-CD16+CD32 antibody [EPR23501-203] (AB223200), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	Flow Cyt	WB	IHC-P	IHC-Fr
Human	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Mouse	Tested	Tested	Tested	Tested	Tested	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Recombinant fragment - Mouse	Not recommended	Not recommended	Not recommended	Tested	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Mouse, Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Mouse, Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Mouse, Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Recombinant fragment - Mouse	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Recombinant fragment - Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Recombinant fragment - Mouse, Human, Rat	Dilution info -	Notes -

Target data

See full target information Fcgr3

Function

Receptor for the Fc region of complexed immunoglobulins gamma. Low affinity receptor which binds to IgG1, IgG2a and IgG2b (PubMed:17558411). Mediates neutrophil activation by IgG complexes redundantly with Fcgr4 (PubMed:18097064).

Additional Targets

Fcgr2

Alternative names

CD16, Low affinity immunoglobulin gamma Fc region receptor III, IgG Fc receptor III, Fc-gamma RIII, FcRIII, Fcgr3

Recommended products

Rabbit Recombinant Monoclonal FCGR3 antibody. Suitable for ICC/IF, IP, Flow Cyt, WB, IHC-P and reacts with Mouse, Recombinant fragment - Mouse samples. Cited in 4 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR23501-203
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CD16 and CD32 also referred to as FCγRIII and FCγRII are proteins expressed on the surface of various immune cells like natural killer cells macrophages and some subsets of T cells. CD16 has a mass ranging between 50-80 kDa while CD32 exhibits variability in size between 32-40 kDa due to different isoforms. These proteins play pivotal roles in immune response by binding to the Fc region of immunoglobulins influencing cell activation and signaling.

Biological function summary

CD16 and CD32 serve important roles in mediating immune cell interactions. They work as part of immune synapses facilitating antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis processes. Due to their involvement in these complex activities they are included in immunological synapse formation and modulation extending their influence over overall immune regulation and response. These proteins are known to contribute to both innate and adaptive immune mechanisms as receptors for the Fc portion of immunoglobulin G.

Pathways

CD16 and CD32 interact within signaling pathways like the Fc gamma receptor-mediated phagocytosis pathway and the immune response-regulating signaling pathway. These receptors work in coordination with other immunoglobulin family members to initiate targeted cellular reactions necessary for immune challenge responses. CD32 often partners with CD19 and CD21 within pathways to modulate B cell receptor-mediated signaling.

Associated diseases and disorders

CD16 and CD32 have connections to autoimmune diseases and infectious diseases. Aberrant expression or dysfunction in these proteins can contribute to the development of conditions such as systemic lupus erythematosus where improper immune complex clearance occurs and chronic inflammation results. CD16's involvement with NK cells can also play a role in viral infection control highlighting their importance in pathogen defense mechanisms.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Flow Cytometry - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Flow cytometric analysis of mouse splenocytes cells labelling CD16+CD32 with ab223200 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (Left).
Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
Flow Cytometry - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Flow cytometric analysis of bEnd.3 (mouse brain endothelioma, Left) / J774A.1 (mouse reticulum cell sarcoma monocyte macrophage, Right) cells labelling CD16+CD32 with ab223200 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Negative control: bEnd.3 (PMID: 23911392).
Gated on viable cells.
Immunoprecipitation - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
CD16+CD32 was immunoprecipitated from 0.35 mg Mouse spleen tissue lysate with ab223200 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab223200 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: Mouse spleen tissue lysate 10 ug
Lane 2: ab223200 IP in Mouse spleen tissue lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab223200 in mouse spleen tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
All lanes: Immunoprecipitation - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Predicted band size: 26 kDa
Observed band size: 40-60 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD16+CD32 with ab223200 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic and membranouse staining on mouse spleen (PMID: 22618994). The section was incubated with ab223200 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunoprecipitation - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
CD16+CD32 was immunoprecipitated from 0.35 mg J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate with ab223200 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab223200 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate 10 ug
Lane 2: ab223200 IP in J774A.1 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab223200 in J774A.1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
All lanes: Immunoprecipitation - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Predicted band size: 26 kDa
Observed band size: 40-60 kDa
Immunocytochemistry/ Immunofluorescence - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) cells labelling CD16+CD32 with ab223200 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing strong membranous and weak cytoplasmic staining in J774A.1 cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Negative control: bEnd.3 cell (PMID: 23911392).
Western blot - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This product can recognize both CD16 and CD32.
The loading samples are E.coil extracts containing CD16 or CD32 recombinant protein respectively.
Exposure time: 70 seconds.
All lanes: Western blot - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution
Lane 1: His-tagged mouse CD16 recombinant protein (aa 31-215), 5 uL
Lane 2: His-tagged mouse CD32 recombinant protein (aa 30-210), 25 uL
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 26 kDa
Western blot - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 19503602, 31172847).
Exposure time: 3 minutes.
All lanes: Western blot - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution
Lane 1: Mouse spleen tissue lysate at 20 µg
Lane 2: Mouse liver tissue lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 26 kDa
Observed band size: 40-60 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Immunohistochemical analysis of paraffin-embedded mouse colon carcinoma tissue labeling CD16+CD32 with ab223200 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on the stroma in mouse colon carcinoma. The section was incubated with ab223200 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling CD16+CD32 with ab223200 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on mouse liver. The section was incubated with ab223200 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
Blocking/Diluting buffer and concentration: 5% NFDM/TBST

The molecular weight observed is consistent with what has been described in the literature (PMID: 19503602, 31172847). Negative control: bEnd.3 (PMID: 23911392).

Exposure time: Lanes 1-2: 5.5 seconds
Lanes 1 - 2: Western blot - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution
Lanes 1 - 2: Western blot - Anti-CD16+CD32 antibody [EPR23501-203] - BSA and Azide free (Anti-CD16+CD32 antibody [EPR23501-203] - BSA and Azide free ab275095)
Lane 1: J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg
Lane 2: bEnd.3 (mouse brain endothelioma) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Observed band size: 40-60 kDa
Exposure time: 5.5s