Recombinant Anti-NKG2A antibody [EPR23737-127] (ab260035)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23737-127] to NKG2A
- Suitable for: Flow Cyt (Intra), IHC-P, mIHC, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-NKG2A antibody [EPR23737-127]
See all NKG2A primary antibodies -
Description
Rabbit monoclonal [EPR23737-127] to NKG2A -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IHC-P, mIHC, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293 transfected with NKG2A (WT) expression vector containing a myc-His-tag, whole cell lysate. IHC-P: Human tonsil, gastric cancer, NK cell lymphoma tissues and HEK-293 transfected with NKG2A expression vector containing a myc-His-tag®. Flow Cyt (intra): Human peripheral blood mononuclear cells. mIHC: Human endometrium tissue.
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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 (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23737-127 -
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 ab260035 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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Flow Cyt (Intra) |
1/500.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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mIHC |
1/2000.
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WB |
1/1000. Predicted molecular weight: 26 kDa.
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Notes |
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Flow Cyt (Intra)
1/500. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
mIHC
1/2000. |
WB
1/1000. Predicted molecular weight: 26 kDa. |
Target
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Function
Plays a role as a receptor for the recognition of MHC class I HLA-E molecules by NK cells and some cytotoxic T-cells. -
Tissue specificity
Natural killer cells. -
Sequence similarities
Contains 1 C-type lectin domain. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 3821 Human
- Omim: 161555 Human
- SwissProt: P26715 Human
- Unigene: 512576 Human
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Alternative names
- C lectin type II protein antibody
- CD159 antigen like family member A antibody
- CD159 antigen-like family member A antibody
see all
Images
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Fluorescence multiplex immunohistochemical analysis of the human endometrium (Formalin/PFA-fixed paraffin-embedded sections).
Panel A: merged staining of anti-EpCAM (ab223582, magenta; Opal™690), anti-NKG2A (ab260035, green; Opal™520) and anti-CD31 (ab182981, red; Opal™570) on human endometrium. Panel B: anti-NKG2A stained on NK cells. Panel C: anti-CD31 stained on endothelial cells. Panel D: anti-EpCAM stained on glandular cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody.
The section was incubated in three rounds of staining: in the order of ab223582 at 1/500 dilution (1.008 μg/ml), ab260035 at 1/2000 dilution (0.262 μg/ml) and ab182981 at 1/4000 dilution (0.137 μg/ml) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of HEK-293 tissue labelling (A) - HEK-293 transfected with NKG2A expression vector containing a myc-His-tag®, (B) - HEK-293 transfected with NKG2C expression vector containing a myc-His-tag®, (C) - HEK-293 transfected with NKG2E expression vector containing a myc-His-tag® whole cell pellets and (D) - HEK-293 transfected with an empty vector with ab260035 at 1/200 dilution. Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins, Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Positive staining on (A) HEK-293 transfected with NKG2A expression vector containing a myc-His-tag®, whole cell pellet. No staining on (B) HEK-293 transfected with NKG2C expression vector containing a myc-His-tag®, whole cell pellet and (C) HEK-293 transfected with NKG2E expression vector containing a myc-His-tag®, whole cell pellet. and (D) HEK-293 transfected with an empty vector.
The section was incubated with ab260035 for30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument -
Immunohistochemical analysis of paraffin-embedded Human NK cell lymphoma tissue labeling NKG2A with ab260035 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human NK cell lymphoma (PMID: 11222501).The section was incubated with ab260035 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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All lanes : Anti-NKG2A antibody [EPR23737-127] (ab260035) at 1/5000 dilution
Lanes 1 & 3 & 5 : HEK-293 (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293 transfected with NKG2A (WT) expression vector containing a myc-His-tag®, whole cell lysate
Lane 4 : HEK-293 transfected with NKG2C (WT) expression vector containing a myc-His-tag®, whole cell lysate
Lane 6 : HEK-293 transfected with NKG2E (WT) expression vector containing a myc-His-tag®, 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
Predicted band size: 26 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
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Intracellular flow cytometric analysis of 2% paraformaldehyde fixed, 0.1% Tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) cells labelling NKG2A with ab260035 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left).
A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Cells were stained with anti-CD56 conjugated to PE, then fixed with 2% PFA followed by intracellularly stained with rabbit IgG (Left) or ab260035 (Right).
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Immunohistochemical analysis of paraffin-embedded Human gastric cancer tissue labeling NKG2A with ab260035 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human gastric cancer (PMID: 11222501).The section was incubated with ab260035 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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Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling NKG2A with ab260035 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human tonsil (PMID: 11222501). The section was incubated with ab260035 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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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab260035 has been referenced in 1 publication.
- Okutani Y et al. Generation of Monkey Induced Pluripotent Stem Cell-Derived Cartilage Lacking Major Histocompatibility Complex Class I Molecules on the Cell Surface. Tissue Eng Part A 28:94-106 (2022). PubMed: 34182799