Recombinant 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

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 (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) 1/200 (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.

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

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

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

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

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

IHC image of ab228462 staining PD-L1 in PD-L1 Dynamic Range Analyte Control formalin fixed paraffin embedded 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.

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

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

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