Anti-NKG2D antibody [RM1173] (ab319162)

Rabbit Recombinant Multiclonal NKG2D antibody. Suitable for IHC-P, Flow Cyt, IP, WB and reacts with Human samples.

Be the first to review this product! Submit a review

Images

Flow Cytometry - Anti-NKG2D antibody [RM1173] (AB319162), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	Flow Cyt	IP	WB	ICC/IF
Human	Tested	Tested	Tested	Tested	Not recommended
Mouse	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	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, Rat	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 Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Target data

See full target information KLRK1

Function

Functions as an activating and costimulatory receptor involved in immunosurveillance upon binding to various cellular stress-inducible ligands displayed at the surface of autologous tumor cells and virus-infected cells. Provides both stimulatory and costimulatory innate immune responses on activated killer (NK) cells, leading to cytotoxic activity. Acts as a costimulatory receptor for T-cell receptor (TCR) in CD8(+) T-cell-mediated adaptive immune responses by amplifying T-cell activation. Stimulates perforin-mediated elimination of ligand-expressing tumor cells. Signaling involves calcium influx, culminating in the expression of TNF-alpha. Participates in NK cell-mediated bone marrow graft rejection. May play a regulatory role in differentiation and survival of NK cells. Binds to ligands belonging to various subfamilies of MHC class I-related glycoproteins including MICA, MICB, RAET1E, RAET1G, RAET1L/ULBP6, ULBP1, ULBP2, ULBP3 (ULBP2>ULBP1>ULBP3) and ULBP4.

Alternative names

CD314, D12S2489E, NKG2D, KLRK1, NKG2-D type II integral membrane protein, Killer cell lectin-like receptor subfamily K member 1, NK cell receptor D, NKG2-D-activating NK receptor

Recommended products

Rabbit Recombinant Multiclonal NKG2D antibody. Suitable for IHC-P, Flow Cyt, IP, WB and reacts with Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: RM1173
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

This product is a recombinant multiclonal 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.

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

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

NKG2D also known as natural killer group 2 member D is a type-2 transmembrane receptor with a mass of approximately 42 kDa. Expressed primarily on natural killer (NK) cells CD8+ T cells subsets of CD4+ T cells and some macrophages it recognizes stress-induced ligands presented on target cells. This protein plays an important role in the immune system's recognition and elimination of infected or transformed cells by binding to its ligands which include MHC class I chain-related proteins MICA/B and UL16-binding proteins.

Biological function summary

NKG2D acts as an activating receptor on the cell surface that initiates immune responses leading to cytotoxicity against target cells. It functions as part of a complex involving the adaptor protein DAP10 which transduces activating signals to promote cellular immune responses. This interaction amplifies the activation of immune cells and contributes to the host's defense against tumors and virally infected cells.

Pathways

NKG2D plays a significant role in immune surveillance and signaling pathways. It actively involves in the natural killer (NK) cell-mediated cytotoxicity pathway and the adaptive immune response pathway. The interaction with proteins such as DAP10 is critical for its signaling function contributing to its interaction within these pathways. The signaling through NKG2D directly affects the activation states and responses of immune cells which are vital in immune defense.

Associated diseases and disorders

NKG2D is implicated in cancer and autoimmune diseases. In cancer overexpression of its ligands on tumor cells makes them susceptible to NKG2D-mediated cell lysis. On the other hand aberrant expression of NKG2D ligands in autoimmune disorders can lead to inappropriate immune activation. The interaction with MICA/B proteins is significant in these conditions emphasizing its importance in both immune surveillance and potential disease pathogenesis.

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.

12 product images

Flow Cytometry - Anti-NKG2D antibody [RM1173] (ab319162)
Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling NKG2D with ab319162 at 1/500 dilution (0.1ug) / right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti human CD8a conjugated to Pacific Blue (440/50BP).

gated on viable lymphocytes.
Flow Cytometry - Anti-NKG2D antibody [RM1173] (ab319162)
Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling NKG2D with ab319162 at 1/500 dilution (0.1ug) / right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti human CD3 conjugated to Alexa fluor 647 and anti human CD56 conjugated to BV421.

Gated on viable CD3 negative cells.
Immunoprecipitation - Anti-NKG2D antibody [RM1173] (ab319162)
NKG2D was immunoprecipitated from 0.35 mg HuT-78 (human sezary syndrome cutaneous t lymphocyte) whole cell lysate with ab319162 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab319162 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HuT-78 (human sezary syndrome cutaneous t lymphocyte) whole cell lysate

Lane 2: ab319162 IP in HuT-78 (human sezary syndrome cutaneous t lymphocyte) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab319162 in HuT-78 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-NKG2D antibody [RM1173] (ab319162) at 1/30 dilution
All lanes: HuT-78 (human sezary syndrome cutaneous t lymphocyte) whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 58s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKG2D antibody [RM1173] (ab319162)
Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling NKG2D with ab319162 at 1/1000 (0.498 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: no staining on human cardiac muscle. The section was incubated with ab319162 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKG2D antibody [RM1173] (ab319162)
Immunohistochemical analysis of paraffin-embedded Human hodgkin's lymphoma tissue labeling NKG2D with ab319162 at 1/1000 (0.498 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human Hodgkin's lymphoma. The section was incubated with ab319162 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKG2D antibody [RM1173] (ab319162)
Immunohistochemical analysis of paraffin-embedded Human non-Hodgkin's lymphoma tissue labeling NKG2D with ab319162 at 1/1000 (0.498 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human non-Hodgkin's lymphoma. The section was incubated with ab319162 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKG2D antibody [RM1173] (ab319162)
Immunohistochemical analysis of paraffin-embedded Human NK/T lymphoma tissue labeling NKG2D with ab319162 at 1/1000 (0.498 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human NK/T lymphoma. The section was incubated with ab319162 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKG2D antibody [RM1173] (ab319162)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling NKG2D with ab319162 at 1/1000 (0.498 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human tonsil. The section was incubated with ab319162 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Western blot - Anti-NKG2D antibody [RM1173] (ab319162)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression tissue: heart
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-NKG2D antibody [RM1173] (ab319162) at 1/1000 dilution
Lane 1: Human tonsil tissue lysate at 20 µg
Lane 2: Human heart tissue lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Observed band size: 35 kDa, 36 kDa
Exposure time: 180s
Western blot - Anti-NKG2D antibody [RM1173] (ab319162)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
NKG2D is a glycoprotein of approximately 35 kDa and detected as a 25-kDa band after treated with N-glycosidase F (PNGase F).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-NKG2D antibody [RM1173] (ab319162) at 1/1000 dilution
Lane 1: Untreated HuT-78 (human Sezary syndrome cutaneous T lymphocyte) whole cell lysate at 15 µg
Lane 2: HuT-78 whole cell lysate treated with N-glycosidase F (PNGase F) at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 35 kDa, 36 kDa
Exposure time: 70s
Western blot - Anti-NKG2D antibody [RM1173] (ab319162)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: Jurkat (PMID: 10426993)
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-NKG2D antibody [RM1173] (ab319162) at 1/1000 dilution
Lane 1: HuT-78 (human Sezary syndrome cutaneous T lymphocyte) whole cell lysate at 20 µg
Lane 2: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 35 kDa, 36 kDa
Exposure time: 180s
Flow Cytometry - Anti-NKG2D antibody [RM1173] (ab319162)
Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling NKG2D with ab319162 at 1/500 dilution (0.1ug) / right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti human CD19 conjugated to Alexa Fluor®647 (670/14BP).

gated on viable lymphocytes.