Anti-NKG2A antibody [EPR23737-127]

• mIHC

Supplier Data

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 (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.

• IHC-P

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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 (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

• IHC-P

Unknown

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 (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.

• IHC-P

Unknown

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 (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.

• Flow Cyt (Intra)

Unknown

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 (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).

• IHC-P

Unknown

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 (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.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-NKG2A antibody [EPR23737-127] (AB260035)

Multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded sections of human NK-T lymphoma tissue labeling CD-161 with ab302564 at 1 : 100 dilution (4.96 μg/mL) colored in red (Opal™570, red). Panel A : merged staining of anti-CD3 (ab16669, grey; Opal™690), anti-CD161(ab302564, red; Opal™570) and anti-NKG2A (ab260035, green; Opal™520) on human NK-T lymphoma. Panel B : anti-CD161 staining only. ab302564 (Opal™ 570, Red) at 1/100 dilution (4.96 µg/mL) Panel C : anti-NKG2A staining only. ; ab260035, (Opal™ 520, Green) NKG2A at 1/2000 dilution (0.283 µg/mL) Panel D : anti-CD3 staining only. CD3 staining only; ab16669 (Opal™ 690, Gray) at 1/2000 dilution (0.0675 µg/mL) The section was incubated in three rounds of staining : in the order of ab16669 for 30 mins, ab302564 for 30 mins and ab260035 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Nuclear stained with DAPI. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used. 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.

• mIHC

Supplier Data

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 (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 (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 ab178089, ab260035, and 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.

• WB

Lab

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 (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 26 kDa

Observed band size: 38 kDa

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