Recombinant Anti-PD-L1 antibody [EPR20529] (ab213480)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20529] to PD-L1
- Suitable for: ICC/IF, IP, WB
- Knockout validated
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-PD-L1 antibody [EPR20529]
See all PD-L1 primary antibodies -
Description
Rabbit monoclonal [EPR20529] to PD-L1 -
Host species
Rabbit -
Specificity
ab213480 is not suitable in IHC-FR for mouse samples
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Tested applications
Suitable for: ICC/IF, IP, WBmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse placenta lysate, HEK mPDL1 cell lysate, whole cell lysate from RAW 264.7 cells treated with Interferon-gamma and whole cell lysate from MEF cells treated with Interferon-gamma. IP: Mouse placenta lysate. ICC/IF: RAW 264.7 cells treated with IFN-? (100 ng/ml) for 24 hours.
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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.
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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20529 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab213480 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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ICC/IF |
Use at an assay dependent concentration.
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IP |
1/30.
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WB | (3) |
1/1000. Detects a band of approximately 40-60 kDa (predicted molecular weight: 33 kDa).
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Notes |
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ICC/IF
Use at an assay dependent concentration. |
IP
1/30. |
WB
1/1000. Detects a band of approximately 40-60 kDa (predicted molecular weight: 33 kDa). |
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: 60533 Mouse
- SwissProt: Q9EP73 Mouse
- Unigene: 245363 Mouse
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Alternative names
- B7 H antibody
- B7 H1 antibody
- B7 homolog 1 antibody
see all
Images
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All lanes : Anti-PD-L1 antibody [EPR20529] (ab213480) at 1/5000 dilution
Lane 1 : Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : RAW264.7 treated with 100 ng/ml IFN gamma for 48 h, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 33 kDa
Observed band size: 40-60 kDa why is the actual band size different from the predicted?
Exposure time: 37 secondsThis WB was performed by using YCA-R20529(BF)-111A H3L2 YR120921PJ, 1:5000 dilution. Working concentration: 0.402 μg/ml.
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
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Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480)This image is courtesy of an anonymous collaborator.All lanes : Anti-PD-L1 antibody [EPR20529] (ab213480) at 100 µg
Lane 1 : HEK mPDL1 cell lysate at 20 µg
Lane 2 : HEK hPDL1 cell lysate at 20 µg
Lane 3 : HEK CL7 at 20 µg
Lane 4 : RAW 264.7 Wild-type (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 100 µg
Lane 5 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate treated with Interferon-gamma at 100 µg
Lane 6 : MEF Wild-type (Mouse embryonic fibroblast cell line) whole cell lysate at 100 µg
Lane 7 : MEF (Mouse embryonic fibroblast cell line) whole cell lysate treated with Interferon-gamma at 100 µg
Lane 8 : MEF mPDL1 KO cell lysate at 100 µg
Lane 9 : MEF mPDL1 KO treated with Interferon-gamma at 100 µg
Predicted band size: 33 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?
Exposure time: 5 minutesThe mAb is specific for mouse mPDL1 and does not recognise hPDL1. As expected a specific 48kDa band is observed in RAW264.7 and MEFS cell extract after Interferon-gamma treatment. No bands are observed in RAW264.7 and MEFS WT and in MEFS KO with and without Interferon-gamma treatment.
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Ab213480 staining PD-L1 in RAW264.7 from Mouse macrophage cell line transformed with Abelson murine leukemia virus by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% Methanol. Samples were incubated with ab213480 at 0.4 μg/ml. AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at 2 μg/ml. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)(ab196889) was used as counterstain at 2.5 μg/ml. DAPI used as nuclear counterstain. Confocal image showing membranous staining on RAW 264.7 cells treated with IFN-γ (100 ng/ml) for 24 hours.
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PD-L1 was immunoprecipitated from 0.35 mg of Mouse placenta lysate with ab213480 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab213480 at 1/500 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution.
Lane 1: Mouse placenta lysate, 10 µg (Input).
Lane 2: ab213480 IP in mouse placenta lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213480 in mouse placenta lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
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Anti-PD-L1 antibody [EPR20529] (ab213480) at 1/1000 dilution + Mouse placenta lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 33 kDa
Observed band size: 40-60 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature. PMID: 15353579.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (44)
ab213480 has been referenced in 44 publications.
- Song X et al. CD169-positive macrophages enhance abscopal effect of radiofrequency ablation therapy in liver cancer. Transl Oncol 15:101306 (2022). PubMed: 34883446
- Moore J et al. Mammary Tumor-Derived Transplants as Breast Cancer Models to Evaluate Tumor-Immune Interactions and Therapeutic Responses. Cancer Res 82:365-376 (2022). PubMed: 34903599
- Mao Z et al. KRAS(G12D) can be targeted by potent inhibitors via formation of salt bridge. Cell Discov 8:5 (2022). PubMed: 35075146
- Wang Y et al. IR792-MCN@ZIF-8-PD-L1 siRNA drug delivery system enhances photothermal immunotherapy for triple-negative breast cancer under near-infrared laser irradiation. J Nanobiotechnology 20:96 (2022). PubMed: 35236356
- Jayathilake AG et al. The comparative anti-cancer effects of krill oil and oxaliplatin in an orthotopic mouse model of colorectal cancer. Nutr Metab (Lond) 19:12 (2022). PubMed: 35236377