Anti-NKG2D 抗体 [RM1173]

• IHC-P

Supplier Data

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

• Flow Cyt

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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 (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488 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.

• IHC-P

Supplier Data

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

• Flow Cyt

Supplier Data

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 (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488 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.

• IHC-P

Supplier Data

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

• IHC-P

Supplier Data

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

• IHC-P

Supplier Data

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

• Flow Cyt

Supplier Data

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 (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488 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.

• IP

Supplier Data

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)(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 (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) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 58s

• WB

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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 (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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 35 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

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 (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

false

Exposure time: 180s

• WB

Supplier Data

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 (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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 35 kDa,36 kDa

false

Exposure time: 70s