Recombinant Anti-CD16+CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)
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 - BSA and Azide free
- Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-CD16+CD32 antibody [EPR23501-203] - BSA and Azide free
See all CD16+CD32 primary antibodies -
Description
Rabbit monoclonal [EPR23501-203] to CD16+CD32 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
Unsuitable for: IHC-Fr -
Species reactivity
Reacts with: Mouse -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: J774A.1 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/IF: 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
ab275095 is the carrier-free version of ab223200.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. -
Storage buffer
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23501-203 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab275095 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 40-60 kDa.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. Detects a band of approximately 40-60 kDa. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Target
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Relevance
Function: Binds to the Fc region of immunoglobulins gamma. Low affinity receptor. By binding to IgG it initiates cellular responses against pathogens and soluble antigens. Promotes phagocytosis of opsonized antigens. Tissue specificity: Found on monocytes, neutrophils and eosinophil platelets. Similarity: Contains 2 Ig-like C2-type (immunoglobulin-like) domains. PTM: Phosphorylated by SRC-type Tyr-kinases such as LYN, BLK, FYN, HCK and SYK. -
Cellular localization
CD32: Type I membrane protein. CD16: Attached to the membrane by a GPI anchor. Exists also as a soluble receptor, produced by a proteolytic cleavage. -
Database links
- Entrez Gene: 14130 Mouse
- Entrez Gene: 14131 Mouse
- SwissProt: P08101 Mouse
- SwissProt: P08508 Mouse
- SwissProt: Q5D5I8 Mouse
- SwissProt: Q5D5J5 Mouse
- SwissProt: Q7TMW9 Mouse
- SwissProt: Q9ES92 Mouse
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Alternative names
- CD16a antibody
- CD16b antibody
- CD32 antibody
see all
Images
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All lanes : Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution
Lane 1 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate
Lane 2 : bEnd.3 (mouse brain endothelioma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Observed band size: 40-60 kDa why is the actual band size different from the predicted?This data was developed using ab223200, the same antibody clone in a different buffer formulation.
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
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This data was developed using ab223200, the same antibody clone in a different buffer formulation.
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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This data was developed using ab223200, the same antibody clone in a different buffer formulation.
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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This data was developed using ab223200, the same antibody clone in a different buffer formulation.
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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This data was developed using ab223200, the same antibody clone in a different buffer formulation.
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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This data was developed using ab223200, the same antibody clone in a different buffer formulation.
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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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 dilutionThis data was developed using ab223200, the same antibody clone in a different buffer formulation.
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.
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This data was developed using ab223200, the same antibody clone in a different buffer formulation.
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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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?This data was developed using ab223200, the same antibody clone in a different buffer formulation.
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.
-
This data was developed using ab223200, the same antibody clone in a different buffer formulation.
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.
-
This data was developed using ab223200, the same antibody clone in a different buffer formulation.
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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab275095 has not yet been referenced specifically in any publications.