United Kingdom (UK)
Recombinant monoclonal antibody to PD-L1. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma JC071.
UniProt Accession Number of Target Protein: E2RKZ5
Alternative Name(s) of Target: CD274; Programmed death ligand 1; PDL-1; PDL1
Immunogen: The original antibody was generated by immunizing BALB/c mice with recombinant canine PD-L1 using an exponentially increasing dosing strategy with MPLA as adjuvant.
Specificity: This antibody binds canine PD-L1.
Application Notes: This antibody is recommended for detection of canine PD-L1. This antibody successfully detected canine PD-L1 in a western blot assay. JC071 was used to determine the increased PD-1 expression on canine PBMCs stimulated with peptidoglycan or lipopolysaccharide using flow cytometry. JC071 was also able to detect recombinant PD-L1 expressed on DH82 cell line both with and without in vitro activation using IFN-γ in flow cytometry. It also recognized recombinant canine PD-L1 expressed on CHO cells in flow cytometry. The ability of the antibody to block interactions between PD-1 and PD-L1 was checked using ELISA. It is known that blockade of PD-L1 and PD-1 interactions causes increase in Ifn-γ level. JC071 successfully blocked PD-L1 and amplified IFN-γ production in culture by approximately 6-fold. Thus, JC071 alleviated T-cell suppression and led to enhanced functional activity of T cells in culture and have potential application as an immunotherapy reagent for cancer treatment (PMID: 32614928).
Antibody first published in:
Choi et al. Development of canine PD-1/PD-L1 specific monoclonal antibodies and amplification of canine T cell function. PLoS One. (2020); 15(7): e0235518 PMID:32614928
Note on publication:
Describes the generation and characterization of antibodies towards canine PD-1 and PD-L1 for their potential use in therapeutics for veterinary oncology.
Western Blot using anti-PD-L1 antibody JC071 (Ab02248). Peripheral blood mononuclear cells lysates (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of JC071 (Ab02248-23.0) at 0.5 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of dog peripheral blood monocytes with anti-PD-L1 (Ab02248) JC071 Immunofluorescence analysis of paraformaldehyde fixed dog peripheral blood monocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of JC071 (Ab02248-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab02248-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.