Anti-NKG2A antibody [EPR23737-127] (ab260035)

Anti-NKG2A antibody [EPR23737-127] (ab260035) is a rabbit monoclonal antibody that is used to detect NKG2A in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, mIHC. Suitable for Human samples.

- Validated for multiplex IHC on the Leica BOND® MAX using Opal reagents

-Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKG2A antibody [EPR23737-127] (AB260035), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 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

	mIHC	WB	Flow Cyt (Intra)	IHC-P
Human	Tested	Expected	Tested	Tested
Mouse	Not recommended	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended
Transfected cell line	Not recommended	Not recommended	Not recommended	Tested
Transfected cell lysate	Not recommended	Tested	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/2000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Transfected cell lysate, Mouse, Rat, Transfected cell line	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Transfected cell lysate	Dilution info 1/1000	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Transfected cell line	Dilution info -	Notes -

Tested
Tested

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

Not recommended
Not recommended

Species	Dilution info	Notes
Species Transfected cell lysate, Mouse, Rat, Transfected cell line	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Transfected cell line	Dilution info 1/2000	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 Mouse	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 Transfected cell lysate	Dilution info -	Notes -

Target data

See full target information KLRC1

Function

Immune inhibitory receptor involved in self-nonself discrimination. In complex with KLRD1 on cytotoxic and regulatory lymphocyte subsets, recognizes non-classical major histocompatibility (MHC) class Ib molecule HLA-E loaded with self-peptides derived from the signal sequence of classical MHC class Ia molecules. Enables cytotoxic cells to monitor the expression of MHC class I molecules in healthy cells and to tolerate self (PubMed:18083576, PubMed:37264229, PubMed:9430220, PubMed:9486650). Upon HLA-E-peptide binding, transmits intracellular signals through two immunoreceptor tyrosine-based inhibition motifs (ITIMs) by recruiting INPP5D/SHP-1 and INPPL1/SHP-2 tyrosine phosphatases to ITIMs, and ultimately opposing signals transmitted by activating receptors through dephosphorylation of proximal signaling molecules (PubMed:12165520, PubMed:9485206). Key inhibitory receptor on natural killer (NK) cells that regulates their activation and effector functions (PubMed:30860984, PubMed:9430220, PubMed:9485206, PubMed:9486650). Dominantly counteracts T cell receptor signaling on a subset of memory/effector CD8-positive T cells as part of an antigen-driven response to avoid autoimmunity (PubMed:12387742). On intraepithelial CD8-positive gamma-delta regulatory T cells triggers TGFB1 secretion, which in turn limits the cytotoxic programming of intraepithelial CD8-positive alpha-beta T cells, distinguishing harmless from pathogenic antigens (PubMed:18064301). In HLA-E-rich tumor microenvironment, acts as an immune inhibitory checkpoint and may contribute to progressive loss of effector functions of NK cells and tumor-specific T cells, a state known as cell exhaustion (PubMed:30503213, PubMed:30860984). (Microbial infection) Viruses like human cytomegalovirus have evolved an escape mechanism whereby virus-induced down-regulation of host MHC class I molecules is coupled to the binding of viral peptides to HLA-E, restoring HLA-E expression and inducing HLA-E-dependent NK cell immune tolerance to infected cells. Recognizes HLA-E in complex with human cytomegalovirus UL40-derived peptide (VMAPRTLIL) and inhibits NK cell cytotoxicity. (Microbial infection) May recognize HLA-E in complex with HIV-1 gag/Capsid protein p24-derived peptide (AISPRTLNA) on infected cells and may inhibit NK cell cytotoxicity, a mechanism that allows HIV-1 to escape immune recognition. (Microbial infection) Upon SARS-CoV-2 infection, may contribute to functional exhaustion of cytotoxic NK cells and CD8-positive T cells (PubMed:32203188, PubMed:32859121). On NK cells, may recognize HLA-E in complex with SARS-CoV-2 S/Spike protein S1-derived peptide (LQPRTFLL) expressed on the surface of lung epithelial cells, inducing NK cell exhaustion and dampening antiviral immune surveillance (PubMed:32859121).

Alternative names

CD159a, NKG2A, KLRC1, NKG2-A/NKG2-B type II integral membrane protein, CD159 antigen-like family member A, NK cell receptor A, NKG2-A/B-activating NK receptor

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 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: EPR23737-127
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

What is this antibody validated in?

Anti-NKG2A antibody [EPR23737-127] (ab260035) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunohistochemistry (IHC-P), Multiplex IHC (mIHC) in Human samples.

What is the molecular weight of NKG2A?

Anti-NKG2A [EPR23737-127] (ab260035) specifically detects a band for NKG2A (UniProt: P26715) at a molecular weight of 26kDa.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products

We have a range of other formats of antibody clone [EPR23737-127] also available for your convenience:
ab260035, Carrier free - ab273516, Alexa Fluor® 647 - ab300745, PE - ab323435

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

What are the advantages of a recombinant monoclonal antibody?
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.

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.

8 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKG2A antibody [EPR23737-127] (ab260035)
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 (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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKG2A antibody [EPR23737-127] (ab260035)
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 (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 (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-NKG2A antibody [EPR23737-127] (ab260035)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
All lanes: Western blot - Anti-NKG2A antibody [EPR23737-127] (ab260035) at 1/5000 dilution
Lanes 1, 3 and 5: HEK-293 (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate at 20 µg
Lane 2: HEK-293 transfected with NKG2A (WT) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg
Lane 4: HEK-293 transfected with NKG2C (WT) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg
Lane 6: HEK-293 transfected with NKG2E (WT) expression vector containing a myc-His-tag®, whole cell 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: 38 kDa
Flow Cytometry (Intracellular) - Anti-NKG2A antibody [EPR23737-127] (ab260035)
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (Left).
A 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.
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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKG2A antibody [EPR23737-127] (ab260035)
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 (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 (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-NKG2A antibody [EPR23737-127] (ab260035)
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 (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 (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.
Multiplex immunohistochemistry - Anti-NKG2A antibody [EPR23737-127] (ab260035)
Fluorescence multiplex immunohistochemical analysis of the human endometrium (Formalin/PFA-fixed paraffin-embedded sections).

Panel A: merged staining of anti-EpCAM (Anti-EpCAM antibody [EPR20532-225] ab223582, magenta; Opal™690), anti-NKG2A (ab260035, green; Opal™520) and anti-CD31 (Anti-CD31 antibody [EPR17259] 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 Anti-EpCAM antibody [EPR20532-225] ab223582 at 1/500 dilution (1.008 μg/ml), ab260035 at 1/2000 dilution (0.262 μg/ml) and Anti-CD31 antibody [EPR17259] 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.
Multiplex immunohistochemistry - Anti-NKG2A antibody [EPR23737-127] (ab260035)
Multichannel (Multiplex) immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections of human tonsil tissue labeling NKG2D, NKG2a, and CD8 alpha.

Panel A: merged staining of anti-CD8 alpha (red; Opal™690), anti-NKG2A (gray; Opal™520) and anti-NKG2D (green; Opal™570) on human tonsil.

Panel B: anti-NKG2D (Anti-NKG2D antibody [EPR24072-342] ab302907 at 1:2000 dilution (0.258 μg/ml)) signal on NK cells and T cell subsets.

Panel C: anti-NKG2A (ab260035 at 1:2000 dilution (0.283 μg/ml)) signal on NK cells.

Panel D: anti-CD8 alpha (Anti-CD8 alpha antibody [SP239] ab178089 at 1:100 dilution (0.83 μg/ml)) signal on T cell subsets.

Opal Polymer HRP Ms + Rb was used as secondary antibody.

The section was incubated in three rounds of staining: in the order of Anti-CD8 alpha antibody [SP239] ab178089, ab260035, and Anti-NKG2D antibody [EPR24072-342] ab302907 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Nuclear counterstain was DAPI.

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.