Recombinant Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) (ab279293)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [CAL10] to PD-L1 - Mouse IgG2a
- Suitable for: ICC/IF, IHC-P, WB, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric)
See all PD-L1 primary antibodies -
Description
Mouse monoclonal [CAL10] to PD-L1 - Mouse IgG2a -
Host species
Mouse -
Tested applications
Suitable for: ICC/IF, IHC-P, WB, Flow Cyt (Intra)more 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
- WB: PD-L1 stably expressed CHO whole cell lysate. Human placenta tissue lysate. NCI-H1299 whole cell lysate. ICC/IF: PD-L1 stably expressed CHO cells. Flow Cyt (intra): PD-L1 stably expressed CHO cells. IHC-P: Human tonsil tissue.
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General notes
This mouse monoclonal chimeric antibody has been engineered from a RabMAb parent antibody (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.
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 (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
CAL10 -
Isotype
IgG2a -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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KO cell lines
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KO cell lysates
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab279293 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 |
1/100.
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IHC-P |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
1/1000.
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Flow Cyt (Intra) |
1/50.
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Notes |
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ICC/IF
1/100. |
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
1/1000. |
Flow Cyt (Intra)
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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All lanes : 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
Lane 2 : Wild-type A549 Vehicle Control IFN-gamma (0 ng/mL, 48 h) cell lysate
Lane 3 : CD274 knockout A549 Treated IFN-gamma (100 ng/mL, 48 h) cell lysate
Lane 4 : CD274 knockout A549 Vehicle Control IFN-gamma (0 ng/mL, 48 h) cell lysate
Lane 5 : Human Placenta cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 45-65 kDa why is the actual band size different from the predicted?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] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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 ab267054 (knockout 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.
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All lanes : 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
Lane 2 : CHO-PD-L1 (PD-L1 stably expressed Chinese hamster ovary epithelial cell) whole cell lysate
Lane 3 : Human placenta tissue lysate
Lane 4 : NCI-H1299 (human lung carcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution
Observed band size: 40-60 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) (ab279293)
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 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
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Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [CAL10] - Mouse IgG2a (Chimeric) (ab279293)
Immunocytochemical analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100-fixed permeabilized CHO-PD-L1 cells labeling PD-L1 with ab279293 at 1/100 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by 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: ab279293 at a 1/100 dilution followed by ab150080 at a 1/200 dilution.
Negative control 2: ab179513 at a 1/200 dilution followed by ab150157 at a 1/1000 dilution.
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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 ab279293 at 1/50 dilution (0.1µg).
Goat Anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
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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SDS download
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Datasheet download
References (0)
ab279293 has not yet been referenced specifically in any publications.