Recombinant Anti-PD-L1 antibody [SP142] - BSA and Azide free (ab236238)
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 - BSA and Azide free
- 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] - BSA and Azide free
See all PD-L1 primary antibodies -
Description
Rabbit monoclonal [SP142] to PD-L1 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, mIHC, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human PD-L1 (C terminal). The exact sequence is proprietary.
Database link: Q9NZQ7 -
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
ab236238 is the carrier-free version of ab228462.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
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. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: 100% PBS -
Carrier free
Yes -
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
- Anti-PD-L1 antibody [SP142] - C-terminal (ab228462)
- Anti-PD-L1 antibody [SP142] - C-terminal, prediluted (ab228463)
- Alexa Fluor® 647 Anti-PD-L1 antibody [SP142] (ab267563)
- Alexa Fluor® 488 Anti-PD-L1 antibody [SP142] - C-terminal (ab310995)
- Alexa Fluor® 568 Anti-PD-L1 antibody [SP142] - C-terminal (ab313030)
- Alexa Fluor® 555 Anti-PD-L1 antibody [SP142] - C-terminal (ab313231)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab236238 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 |
Use at an assay dependent concentration.
Boil tissue section in EDTA buffer, pH 8.0 for 10 minutes followed by cooling at room temperature for 20 minutes. Primary antibody incubation for 10 minutes at room temperature.
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mIHC |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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IHC-P
Use at an assay dependent concentration. Boil tissue section in EDTA buffer, pH 8.0 for 10 minutes followed by cooling at room temperature for 20 minutes. Primary antibody incubation for 10 minutes at room temperature.
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mIHC
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
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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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. -
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).
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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.
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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.
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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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
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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.
This image was generated using ab228462, the same antibody but with BSA and Azide
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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.
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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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).
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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.
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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Formalin-fixed, paraffin-embedded human prostate adenocarcinoma 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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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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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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (2)
ab236238 has been referenced in 2 publications.
- Altorki NK et al. Global evolution of the tumor microenvironment associated with progression from preinvasive invasive to invasive human lung adenocarcinoma. Cell Rep 39:110639 (2022). PubMed: 35385730
- McDowell SAC et al. Neutrophil oxidative stress mediates obesity-associated vascular dysfunction and metastatic transmigration. Nat Cancer 2:545-562 (2021). PubMed: 35122017