Recombinant Anti-PD-L1 antibody [SP142] - C-terminal (ab228462)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP142] to PD-L1 - C-terminal
- Suitable for: IHC-P, mIHC, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-PD-L1 antibody [SP142] - C-terminal
See all PD-L1 primary antibodies -
Description
Rabbit monoclonal [SP142] to PD-L1 - C-terminal -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, mIHC, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human placenta, tonsil, lung squamous cell carcinoma, cervical squamous cell carcinoma, skin squamous cell carcinoma, Hodgkin's lymphoma, pancreatic adenocarcinoma and prostate adenocarcinoma tissues. ICC/IF: CHO-PD-L1 cells. mIHC: Human tonsil
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading...
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Purity
Protein A/G purified -
Purification notes
Purified from TCS by protein A/G. -
Clonality
Monoclonal -
Clone number
SP142 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab228462 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P | (2) |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Primary antibody incubation for 10 minutes at room temperature. |
mIHC |
1/500.
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ICC/IF |
1/50.
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Notes |
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IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Primary antibody incubation for 10 minutes at room temperature. |
mIHC
1/500. |
ICC/IF
1/50. |
Target
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Function
Involved in the costimulatory signal, essential for T-cell proliferation and production of IL10 and IFNG, in an IL2-dependent and a PDCD1-independent manner. Interaction with PDCD1 inhibits T-cell proliferation and cytokine production. -
Tissue specificity
Highly expressed in the heart, skeletal muscle, placenta and lung. Weakly expressed in the thymus, spleen, kidney and liver. Expressed on activated T- and B-cells, dendritic cells, keratinocytes and monocytes. -
Sequence similarities
Belongs to the immunoglobulin superfamily. BTN/MOG family.
Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
Cellular localization
Cell membrane and Endomembrane system. - Information by UniProt
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Database links
- Entrez Gene: 29126 Human
- Omim: 605402 Human
- SwissProt: Q9NZQ7 Human
- Unigene: 521989 Human
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Alternative names
- B7 H antibody
- B7 H1 antibody
- B7 homolog 1 antibody
see all
Images
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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
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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 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
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
Datasheets and documents
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SDS download
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Datasheet download
References (87)
ab228462 has been referenced in 87 publications.
- Song JY et al. Low T-cell proportion in the tumor microenvironment is associated with immune escape and poor survival in diffuse large B-cell lymphoma. Haematologica N/A:N/A (2023). PubMed: 36632739
- Sakatani T et al. IFN-Gamma Expression in the Tumor Microenvironment and CD8-Positive Tumor-Infiltrating Lymphocytes as Prognostic Markers in Urothelial Cancer Patients Receiving Pembrolizumab. Cancers (Basel) 14:N/A (2022). PubMed: 35053427
- Wu SY et al. Combined angiogenesis and PD-1 inhibition for immunomodulatory TNBC: concept exploration and biomarker analysis in the FUTURE-C-Plus trial. Mol Cancer 21:84 (2022). PubMed: 35337339
- Hu L et al. Clinical relevance of pathogenic germline variants in mismatch repair genes in Chinese breast cancer patients. NPJ Breast Cancer 8:52 (2022). PubMed: 35449176
- Li Z et al. Mutual exclusivity of ESR1 and TP53 mutations in endocrine resistant metastatic breast cancer. NPJ Breast Cancer 8:62 (2022). PubMed: 35538119