Knockout Tested Mouse Recombinant Monoclonal PD-L1 antibody. Carrier free. Suitable for IHC-P, ICC, WB, Flow Cyt (Intra) and reacts with Human, Recombinant fragment - Human samples.
IgG2a
Mouse
Constituents: 100% PBS
Liquid
Monoclonal
IHC-P | ICC | WB | Flow Cyt (Intra) | |
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Human | Tested | Expected | Tested | Expected |
Recombinant fragment - Human | Not recommended | Tested | Tested | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Recombinant fragment - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Recombinant fragment - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Recombinant fragment - Human, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Recombinant fragment - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
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Plays a critical role in induction and maintenance of immune tolerance to self (PubMed:11015443, PubMed:28813410, PubMed:28813417, PubMed:31399419). As a ligand for the inhibitory receptor PDCD1/PD-1, modulates the activation threshold of T-cells and limits T-cell effector response (PubMed:11015443, PubMed:28813410, PubMed:28813417, PubMed:36727298). Through a yet unknown activating receptor, may costimulate T-cell subsets that predominantly produce interleukin-10 (IL10) (PubMed:10581077). Can also act as a transcription coactivator: in response to hypoxia, translocates into the nucleus via its interaction with phosphorylated STAT3 and promotes transcription of GSDMC, leading to pyroptosis (PubMed:32929201).The PDCD1-mediated inhibitory pathway is exploited by tumors to attenuate anti-tumor immunity and escape destruction by the immune system, thereby facilitating tumor survival (PubMed:28813410, PubMed:28813417). The interaction with PDCD1/PD-1 inhibits cytotoxic T lymphocytes (CTLs) effector function (By similarity). The blockage of the PDCD1-mediated pathway results in the reversal of the exhausted T-cell phenotype and the normalization of the anti-tumor response, providing a rationale for cancer immunotherapy (By similarity).
B7H1, PDCD1L1, PDCD1LG1, PDL1, CD274, Programmed cell death 1 ligand 1, PD-L1, PDCD1 ligand 1, Programmed death ligand 1, hPD-L1, B7 homolog 1, B7-H1
Knockout Tested Mouse Recombinant Monoclonal PD-L1 antibody. Carrier free. Suitable for IHC-P, ICC, WB, Flow Cyt (Intra) and reacts with Human, Recombinant fragment - Human samples.
IgG2a
Mouse
Constituents: 100% PBS
Liquid
Monoclonal
Yes
CAL10
Affinity purification Protein A
Blue Ice
+4°C
+4°C
Do Not Freeze
ab279305 is the carrier free version of Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293.
This mouse monoclonal chimeric antibody has been engineered from a RabMAb parent antibody (Anti-PD-L1 antibody [CAL10] ab237726). By necessity, some rabbit sequence is retained as part of the variable domain. When multiplexing with other rabbit-derived antibodies, using cross absorbed Fc-reactive secondary antibodies are recommended.
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 supplementary information is collated from multiple sources and compiled automatically.
PD-L1 also known as Programmed Death-Ligand 1 or CD274 is a protein involved in immune modulation. Mechanically PD-L1 interacts with its receptors particularly PD-1 to regulate cellular immune responses. This transmembrane protein has a calculated molecular weight of approximately 33 kDa. PD-L1 is expressed on various cell types including tumor cells and immune cells such as dendritic cells macrophages and B cells. Its expression is often upregulated in response to inflammatory cytokines.
PD-L1 plays a central role in immune evasion mechanisms utilized by tumors. It is not part of a larger protein complex but directly interacts with PD-1 and CD80. When PD-L1 binds to PD-1 it sends inhibitory signals leading to decreased T cell activation and proliferation allowing cancer cells to avoid immune destruction. PD-L1 expression provides a mechanism for tumors to suppress immune surveillance facilitating tumor progression.
PD-L1 is integral to the immune checkpoint pathway which is an important regulator of immune response. The interaction between PD-L1 and PD-1 provides a mechanism for immune tolerance which is part of the broader adaptive immune system pathway. PD-L1 is related to other immune checkpoint proteins such as CTLA-4 in its function to limit autoreactivity and promote immune homeostasis.
PD-L1 is most associated with cancer particularly in tumors such as melanoma and non-small cell lung cancer. PD-L1 expression on tumor cells often correlates with poor prognosis. PD-L1 directly interacts with PD-1 in these cancers a target for immunotherapies such as checkpoint inhibitors which aim to block this interaction and restore immune activity against tumors. PD-L1 involvement extends to autoimmune disorders where altered expression can impact tolerance and lead to immune-related tissue damage.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
This data was produced using Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293, the same clone in a different formulation.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-PD-L1 antibody [CAL10] – Mouse IgG2a (Chimeric) (Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293) at 1/1000 dilution
Lane 1: CHO-S (Chinese hamster ovary epithelial cell) whole cell lysate at 20 µg
Lane 2: CHO-PD-L1 (PD-L1 stably expressed Chinese hamster ovary epithelial cell) whole cell lysate at 20 µg
Lane 3: Human placenta tissue lysate at 20 µg
Lane 4: NCI-H1299 (human lung carcinoma epithelial cell), whole cell lysate at 20 µg
All lanes: Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution
Predicted band size: 33 kDa
Observed band size: 40-60 kDa
This data was produced using Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293, the same clone in a different formulation.
IHC image of PD-L1 staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BONDTM system using the standard protocol F.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293, 1ug/ml, for 15 mins at room temperature. A rabbit anti-mouse IgG2a, was added for 8 mins at room temperature and detected using an HRP conjugated goat anti-rabbit compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
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
This data was produced using Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293, the same clone in a different formulation.
Immunocytochemical analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100-fixed permeabilized CHO-PD-L1 cells labeling PD-L1 with Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/500 dilution (Red). The nuclear counterstain was DAPI (Blue). Confocal image showing membranous and cytoplasmic staining in CHO-PD-L1 cells.
Negative control 1: Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293 at a 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 at a 1/200 dilution.
Negative control 2: Anti-beta Tubulin antibody [EPR16774] ab179513 at a 1/200 dilution followed by Goat Anti-Rat IgG H&L (Alexa Fluor® 488) ab150157 at a 1/1000 dilution.
This data was produced using Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293, the same clone in a different formulation.
Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized CHO-s (Chinese hamster ovary epithelial cell, Blue) / CHO-PDL1 (PD-L1 stably expessed Chinese hamster ovary epithelial cell, Red) labelling PD-L1 with Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293 at 1/50 dilution (0.1μg).
Goat Anti-Mouse IgG (Alexa Fluor® 488, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/2000 dilution was used as the secondary antibody.
False colour image of Western blot: Anti-PD-L1 antibody [CAL10] – Mouse IgG2a (Chimeric) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293 was shown to bind specifically to PD-L1. A band was observed at 45-65 kDa in treated wild-type A549 cell lysates with no signal observed at this size in Cd274 knockout cell line Human CD274 (PD-L1) knockout A549 cell line ab267054 (knockout cell lysate Human CD274 (PD-L1) knockout A549 cell lysate ab256831). To generate this image, wild-type and Cd274 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) (Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) ab279293) at 1/1000 dilution
Lane 1: Wild-type A549 Treated IFN-gamma (100 ng/mL, 48 h) cell lysate at 20 µg
Lane 2: Wild-type A549 Vehicle Control IFN-gamma (0 ng/mL, 48 h) cell lysate at 20 µg
Lane 3: CD274 knockout A549 Treated IFN-gamma (100 ng/mL, 48 h) cell lysate at 20 µg
Lane 4: CD274 knockout A549 Vehicle Control IFN-gamma (0 ng/mL, 48 h) cell lysate at 20 µg
Lane 5: Human Placenta cell lysate at 20 µg
Lanes 1 - 5: Goat anti-Mouse IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 5: Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 45-65 kDa
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