Anti-CD161 抗体 [EPR26340-6] (ab302564)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling CD161 with ab302564 at 1/100 dilution (4.96 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). Positive staining on human spleen. The section was incubated with ab302564 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling CD161 with ab302564 at 1/100 dilution (4.96 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). Positive staining on immune cells of human endometrium. The section was incubated with ab302564 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling CD161 with ab302564 at 1/100 dilution (4.96 µg/mL) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). Positive staining on immune cells of human colon. The section was incubated with ab302564 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a Myc-His tagged CD161 construct and (B) HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with empty plasmid labeling CD161 with ab302564 at 1/100 dilution (4.96 µg/mL) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). Positive staining on (A) HEK-293T transfected with a Myc-His tagged CD161 construct, no staining on (B) HEK-293T transfected with empty plasmid. The section was incubated with ab302564 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling CD161 with ab302564 at 1/100 dilution (4.96 µg/mL) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). Negative control: no staining on human skeletal muscle. The section was incubated with ab302564 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

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.

Multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded sections of human tonsil tissue labeling CD-161 with ab302564 at 1/100 dilution (4.96 μg/mL), NKG2A with ab260035 at 1/2000 dilution ( 0.283 μg/ml) and CD3 with ab16669 at 1/2000 dilution (0.0675 µg/mL).

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.
Panel C: anti-NKG2A staining only.
Panel D: anti-CD3 staining only.

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.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized human PBMC (human peripheral blood mononuclear cell) cells labeling CD161 with AB302564 at 1/50 (9.92 µg/mL) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preabsorbed antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing membrane and cytoplasmic staining in subsets of human PBMC. Ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody control: Primary diluent was used instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preabsorbed at 1/1000 dilution (2 µg/mL).

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labeling CD161 with AB302564 at 1/500 dilution (0.1 µg)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Cells were co-stained with anti-CD4 conjugated to APC/Fire™ 750. Gated on viable cells.

Flow cytometric analysis of human PBMC (human peripheral blood mononuclear cell) cells labeling CD161 with AB302564 at 1/500 dilution (0.1 µg)/ Right compared with a rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: B lymphocytes (CD19⁺). Gated on viable cells.