Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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.

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

Clone SP142 (ab236238) has been successfully conjugated by Abcam. This image was generated using Anti-PD-L1 antibody [SP142] (Alexa Fluor® 647). Please refer to ab267563 for protocol details.

IHC image of PD-L1 staining in a section of formalin-fixed paraffin-embedded normal human placenta*.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Biocare Medical NxGen pressure cooker using retrieval settings of 110°C for 20 minutes. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab267563 at 1/100 dilution (shown in red) and counterstained using ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

*Tissue obtained from Papworth Hospital Research Tissue Bank.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228462).

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling PD-L1 with ab236238 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human placenta, performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with EDTA-Tris buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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.

This image was generated using ab228462, the same antibody but with BSA and Azide

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228462).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228462).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228462).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228462).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228462).

• mIHC

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Multiplex immunohistochemistry - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

This data was developed using ab228462, the same antibody clone in a different buffer formulation. 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.

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228462).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228462).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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.

This image was generated using ab228462, the same antibody but with BSA and Azide

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [SP142] - C-terminal, BSA and Azide free (AB236238)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228462).