Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
![Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-6-anti-cd16cd32-antibody-epr23501203-flow-cytometry-mouse-splenocytes.jpg "Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23501-203] to CD16+CD32
- Suitable for: Flow Cyt, IP, ICC, WB, IHC-P
- Reacts with: Mouse, Recombinant fragment
Overview
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Product name
Anti-CD16+CD32 antibody [EPR23501-203]
See all CD16+CD32 primary antibodies -
Description
Rabbit monoclonal [EPR23501-203] to CD16+CD32 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt MouseICC MouseIHC-P MouseIP MouseWB MouseRecombinant fragment
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Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: J774A.1 and DC2.4 whole cell lysates; Mouse spleen and liver tissue lysates; His-tagged mouse CD16 recombinant protein; His-tagged mouse CD32 recombinant protein. IHC-P: Mouse spleen, liver and colon carcinoma tissue. ICC: J774A.1 cells. Flow cyt: J774A.1 and mouse splenocyte cells. IP: J774A.1 whole cell lysate; Mouse spleen tissue lysate.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23501-203 -
Isotype
IgG -
Research areas
Images
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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 (ab172730) isotype control (Left).
Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
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![Flow Cytometry - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-5-anti-cd16cd32-antibody-epr23501203-flow-cytometry-bend3.jpg)
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 (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, ab150077) at 1/2000 dilution was used as the secondary antibody.
Negative control: bEnd.3 (PMID: 23911392).
Gated on viable cells.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-1-anti-cd16cd32-antibody-epr23501203-immunohistochemistry-mouse-spleen.jpg)
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 (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 (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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![Immunocytochemistry - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-4-anti-cd16cd32-antibody-epr23501203-immunocytochemistry-j774a1.jpg)
Immunocytochemistry - 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 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. 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Negative control: bEnd.3 cell (PMID: 23911392).
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![Immunoprecipitation - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-8-anti-cd16cd32-antibody-epr23501203-immunoprecipitation-mouse-spleen-tissue-lysate-10-ug.jpg)
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)(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 (ab172730) instead of ab223200 in mouse spleen tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-411853-anti-cd16cd32-antibody-epr23501203-western-blot-recombinant.jpg)
Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)All lanes : 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 (ab97051) at 1/50000 dilutionBlocking 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.
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![Immunoprecipitation - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-7-anti-cd16cd32-antibody-epr23501203-immunoprecipitation-j774a1.jpg)
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)(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 (ab172730) instead of ab223200 in J774A.1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
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![Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-411852-anti-cd16cd32-antibody-epr23501203-western-blot-lysate.jpg)
Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)All lanes : Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution
Lane 1 : Mouse spleen tissue lysate
Lane 2 : Mouse liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Observed band size: 40-60 kDa why is the actual band size different from the predicted?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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-3-anti-cd16cd32-antibody-epr23501203-immunohistochemistry-mouse-colon-carcinoma.jpg)
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 (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 (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)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-411851-anti-cd16cd32-antibody-epr23501203-western-blot-cell.jpg)
Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)All lanes : Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution
Lane 1 : J774A.1 (mouse reticum cell sarcoma monocyte macrophage) whole cell lysate
Lane 2 : bEnd.3 (mouse brain endothelioma) whole cell lysate
Lane 3 : DC2.4 (mouse dendritic cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Observed band size: 40-60 kDa why is the actual band size different from the predicted?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).
Negative control: bEnd.3 (PMID: 23911392).
Exposure time: Lanes 1-2: 5.5 seconds; Lane 3: 59 seconds.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-2-anti-cd16cd32-antibody-epr23501203-immunohistochemistry-mouse-liver.jpg)
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 (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 (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
![Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)](https://www.abcam.com/ps/products/223/ab223200/Images/ab223200-12-benefits-of-recombinant-antibodies.png)
Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)