Anti-PD-L1 抗体 [SP142] - C-terminal

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Formalin-fixed, paraffin-embedded human lung squamous cell carcinoma tissue stained for PD-L1 using ab228462 at 1/100 dilution in immunohistochemical analysis.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• mIHC

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Multiplex immunohistochemistry - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining PD1 with ab309363 at a 1/4000 dilution ( 0.125 μg/ml), ab228462 anti-PD-L1 used at 1/100 dilution (0.52 μg/ml) and ab213363 anti-CD68 used at a 1/500 dilution (1.26 μg/ml).

Panel A : merged staining of anti-CD68 (gray; Opal™690), anti-PD1 (green; Opal™520) and anti-PD-L1 (red; Opal™570) on human tonsil.
Panel B : anti-PD1 stained on antigen-stimulated T cells.
Panel C : anti-PD-L1 stained on cells involved in T cell inhibition.
Panel D : anti-CD68 stained on macrophages.

The section was incubated in three rounds of staining : in the order of ab213363 and ab309363 for 30 mins, then ab228462 for 10 mins at room temperature.

Heat mediated antigen retrieval 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Formalin-fixed, paraffin-embedded human skin squamous cell carcinoma tissue stained for PD-L1 using ab228462 at 1/100 dilution in immunohistochemical analysis.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

IHC image of ab228462 staining PD-L1 in PD-L1 Dynamic Range Analyte Control formalin fixed paraffin embedded human cell lines (HistoCyte Laboratories), performed on a Leica BOND RX (standard Protocol F, Polymer Refine kit). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH9, epitope retrieval solution 2) for 30 mins at 98°C. The section was then incubated with ab228462, 1/400 working dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system for 8 minutes at room temperature. DAB was used as the chromogen for 10 minutes at room temperature. The section was then counterstained with hematoxylin, blued, dehydrated, cleared and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Formalin-fixed, paraffin-embedded human Hodgkin's lymphoma tissue stained for PD-L1 using ab228462 at 1/100 dilution in immunohistochemical analysis.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Formalin-fixed paraffin-embedded human pancreatic adenocarcinoma tissue stained for PD-L1 using ab228462 at 1/100 dilution in immunohistochemical analysis.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Formalin-fixed paraffin-embedded human prostate adenocarcinoma tissue stained for PD-L1 using ab228462 at 1/100 dilution in immunohistochemical analysis.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Formalin-fixed paraffin-embedded human placenta tissue stained for PD-L1 using ab228462 at 1/100 dilution in immunohistochemical analysis.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Formalin-fixed paraffin-embedded human tonsil tissue stained for PD-L1 using ab228462 at 1/100 dilution in immunohistochemical analysis.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling PD1 with ab243644 at 1/500 dilution (1.02 µg/mL) (D), Ki67 with ab16667 at 1/200 dilution (0.15 μg/ml) (B) and PD-L1 with ab228462 at 1/100 dilution (0.52 μg/ml) (C). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Panel A : merged staining of anti-Ki67 (magenta; Opal™690), anti-PD-L1 (red; Opal™570) and anti-PD1 (green; Opal™520) on human tonsil. Panel B : anti-Ki67 stained on nucleus of proliferating cells. Panel C : anti-PD-L1 stained on membrane of cells involved in T cell inhibition. Panel D : anti-PD1 stained on antigen-stimulated T cells. The section was incubated in three rounds of staining : in the order of ab16667 for 10 mins, ab243644 for 30 mins and ab228462 for 10 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Formalin-fixed, paraffin-embedded human cervical squamous cell carcinoma tissue stained for PD-L1 using ab228462 at 1/100 dilution in immunohistochemical analysis.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Immunohistochemical analysis of formalin fixed paraffin (FFPE) embedded tonsil labelling PD-L1 with ab228462 at a dilution of 1/200. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit followed by OptiView Amplification kit . Heat mediated antigen retrieval was conducted for 32min with DISCOVERY cell conditioning solution (CC1)  100°C, pH 8.5. ab228462 was incubated at 37°C for 16 min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

IHC image of ab228462 staining PD-L1 in human tonsil formalin fixed paraffin embedded tissue sections*, performed on a Leica BOND RX (standard Protocol F, Polymer Refine kit). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH9, epitope retrieval solution 2) for 30 mins at 98°C. The section was then incubated with ab228462, 1/400 working dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system for 8 minutes at room temperature. DAB was used as the chromogen for 10 minutes at room temperature. The section was then counterstained with hematoxylin, blued, dehydrated, cleared and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Tissue Microarrays stained for "Anti-PD-L1 antibody [SP142] - C-terminal" using "ab228462"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab228462 for 10 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [SP142] - C-terminal (AB228462)

Immunocytochemistry/ Immunofluorescence analysis of CHO-PD-L1 (PD-L1 stably expessed Chinese hamster ovary epithelial cell) cells labeling PD-L1 with purified ab228462 at 1/50 dilution (2 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488 ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.