Anti-NKR-P1C antibody [EPR22990-31]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKR-P1C antibody [EPR22990-31] (AB289542)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling NKR-P1C with ab289542 at 1/100 (5.26 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on mouse spleen. The section was incubated with ab289542 at 4°C overnight. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NKR-P1C antibody [EPR22990-31] (AB289542)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse spleen cells labelling NKR-P1C with ab289542 at 1/50 (10.52 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 μg/ml) dilution (Green). Confocal image showing membranal staining in subsets of mouse splenocytes is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 μg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 μg/ml) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKR-P1C antibody [EPR22990-31] (AB289542)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling NKR-P1C with ab289542 at 1/100 (5.26 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on lympho node of mouse colon. The section was incubated with ab289542 at 4°C overnight. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• Flow Cyt

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Flow Cytometry - Anti-NKR-P1C antibody [EPR22990-31] (AB289542)

Flow cytometric analysis of Mouse splenocytes cells labelling NKR-P1C with ab289542 at 1/500 dilution (0.1ug). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Cells were stained with ab289542, and then stained with anti-CD19 conjugated to PE-Cy7 or anti-CD11b conjugated to BV510. Gated on viable cells. Negative expression on B cells (Left) and Myeloid cells (Right) were observed.

• Flow Cyt

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Flow Cytometry - Anti-NKR-P1C antibody [EPR22990-31] (AB289542)

Flow cytometric analysis of Mouse splenocytes cells labelling NKR-P1C with ab289542 at 1/500 dilution (0.1ug) (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab289542. Then stained with anti-CD49b conjugated to PE. Gated on viable cells.

• Flow Cyt

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Flow Cytometry - Anti-NKR-P1C antibody [EPR22990-31] (AB289542)

Flow cytometric analysis of Mouse splenocytes cells labelling NKR-P1C with ab289542 at 1/500 dilution (0.1ug). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Cells were stained with ab289542, and then stained with anti-CD3 conjugated to Alexa Fluor® 647. Gated on viable cells. NK(NKR-P1C+) and NKT(CD3+NKR-P1C+) population were observed by CD3 co-staining.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-NKR-P1C antibody [EPR22990-31] (AB289542)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen (fresh) tissue labeling NKR-P1C with ab289542 at 1/100 (5.26 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 μg/mL) dilution (Green). Positive staining in mouse spleen is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 μg/mL) dilution.

• IHC-P

AbReview82632****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKR-P1C antibody [EPR22990-31] (AB289542)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of paraformaldehyde-fixed mouse lung tissue staining with ab289542 at 1/100 dilution. Secondary antibody was a ready to use HRP anti-polyvalent. Samples were incubated with the primary antibody with 1% BSA in 1x PBS for 24 hours at 4°C. Blocking was done using a ready to use 12.5% Mouse on Mouse Blocking reagent for 1 hour at 22 degrees. Heat mediated antigen retrieval with Tris/EDTA pH9 performed in a steamer at approx. 99°C for 20 min. Hematoxylin was used for counterstaining.

This image is courtesy of an anonymous Abreview

• mIHC

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Multiplex immunohistochemistry - Anti-NKR-P1C antibody [EPR22990-31] (AB289542)

Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse spleen tissue staining Klra8 with ab313388 at a 1/2000 dilution ( 0.253 μg/ml), ab289542 anti-NKR-P1C used at 1/100 dilution (5.18 μg/ml). Panel A : merged staining of anti-NKR-P1C (green; Opal™520) and anti-Klra8 (red; Opal™570) on mouse spleen. Panel B : anti-Klra8 stained on NK cell subsets. Panel C : anti-NKR-P1C stained on NK cell subsets. The section was incubated in two rounds of staining : in the order of ab289542 and ab313388 for 30 mins at room temperature. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins 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.