Recombinant Anti-CD16 antibody [EPR22409-124] (ab246222)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22409-124] to CD16
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD16 antibody [EPR22409-124]
See all CD16 primary antibodies -
Description
Rabbit monoclonal [EPR22409-124] to CD16 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human spleen and colon cancer lysates. IP: Human spleen lysate. IHC-P: Human spleen and liver tissue. ICC/IF: Human PBMCs. Flow Cyt: Human PBMCs.
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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.
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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22409-124 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab246222 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 |
1/1000. Detects a band of approximately 50-70 kDa (predicted molecular weight: 29 kDa).
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IHC-P | (1) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/100.
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Flow Cyt |
1/600.
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 50-70 kDa (predicted molecular weight: 29 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
Flow Cyt
1/600. |
IP
1/30. |
Target
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Function
Receptor for the Fc region of IgG. Binds complexed or aggregated IgG and also monomeric IgG. Mediates antibody-dependent cellular cytotoxicity (ADCC) and other antibody-dependent responses, such as phagocytosis. -
Tissue specificity
Expressed on natural killer cells, macrophages, subpopulation of T-cells, immature thymocytes and placental trophoblasts. -
Sequence similarities
Contains 2 Ig-like C2-type (immunoglobulin-like) domains. -
Post-translational
modificationsGlycosylated. Contains high mannose- and complex-type oligosaccharides.
The soluble form is produced by a proteolytic cleavage. -
Cellular localization
Cell membrane. Secreted. Exists also as a soluble receptor. - Information by UniProt
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Database links
- Entrez Gene: 2214 Human
- Omim: 146740 Human
- SwissProt: P08637 Human
- Unigene: 372679 Human
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Alternative names
- CD 16 antibody
- CD 16a antibody
- CD16 antibody
see all
Images
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CD16 was immunoprecipitated from 0.35 mg of human spleen lysate with ab246222 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab246222 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as the secondary antibody at 1/5000 dilution.
Lane 1: Human spleen lysate 10 µg (Input).
Lane 2: ab246222 IP in human spleen lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab246222 in human spleen lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds. -
Human PBMCs (human peripheral blood mononuclear cells) were stained with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left) or ab246222 (Right) followed by a Goat anti rabbit IgG (Dylight ® 488) at 1/2000 dilution. They were then stained with Alexa Fluor® 647-conjugated anti-CD14. The expression pattern is consistent with that described in the literature. (PMID: 21738687)
Gated on viable cells.
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Negative control: Raji. (PMID:11207281).
Raji (human Burkitt's lymphoma cell line) were stained with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)(Left) or ab246222 (Right) followed by a Goat anti rabbit IgG (Dylight ® 488) at 1/2000 dilution.
Gated on viable cells.
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Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD16 with ab246222 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Kuffer cells of human liver (PMID: 26512139) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling CD16 with ab246222 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on red pulp of human spleen (PMID: 26512139; 29692344). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized human PBMCs (human peripheral blood mononuclear cells) or Raji (human Burkitt's lymphoma cell line) cell line labeling CD16 with ab246222 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in subsets of human PBMC cells is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
Negative control: Raji (PMID:11207281).
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All lanes : Anti-CD16 antibody [EPR22409-124] (ab246222) at 1/1000 dilution
Lane 1 : Human spleen lysate
Lane 2 : Human colon cancer lysate
Lane 3 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/100000 dilution
Predicted band size: 29 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times.
Lane 1: 92 seconds.
Lanes 2 & 3: 3 minutes.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 2526846).
Negative control: Raji (PMID:11207281).
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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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab246222 has been referenced in 3 publications.
- Tan Y et al. RNA interference-mediated silencing of DNA methyltransferase 1 attenuates neuropathic pain by accelerating microglia M2 polarization. BMC Neurol 22:376 (2022). PubMed: 36183073
- Zheng G et al. Interaction between HLA-G and NK cell receptor KIR2DL4 orchestrates HER2-positive breast cancer resistance to trastuzumab. Signal Transduct Target Ther 6:236 (2021). PubMed: 34158475
- Zhang P et al. Decreased IL-6 and NK Cells in Early-Stage Lung Adenocarcinoma Presenting as Ground-Glass Opacity. Front Oncol 11:705888 (2021). PubMed: 34568032