About RecombiMAb anti-mouse PD-1 (CD279) (D265A) The 29F.1A12™-CP005 monoclonal antibody is a recombinant chimeric version of the original 29F.1A12™ antibody. The variable domain sequences are identical to the original 29F.1A12™ but the constant region sequences have been switched from rat IgG2a to mouse IgG1. The 29F.1A12™-CP005 antibody also contains a D265A mutation in the Fc fragment rendering it unable to bind to endogenous Fcγ receptors. 29F.1A12™-CP005 reacts with mouse PD-1 (programmed death-1) also known as CD279. PD-1 is a 50-55 kDa cell surface receptor encoded by the Pdcd1 gene that belongs to the CD28 family of the Ig superfamily. PD-1 is transiently expressed on CD4 and CD8 thymocytes as well as activated T and B lymphocytes and myeloid cells. PD-1 expression declines after successful elimination of antigen. Additionally, Pdcd1 mRNA is expressed in developing B lymphocytes during the pro-B-cell stage. PD-1’s structure includes a ITIM (immunoreceptor tyrosine-based inhibitory motif) suggesting that PD-1 negatively regulates TCR signals. PD-1 signals via binding its two ligands, PD-L1 and PD-L2 both members of the B7 family. Upon ligand binding, PD-1 signaling inhibits T-cell activation, leading to reduced proliferation, cytokine production, and T-cell death. Additionally, PD-1 is known to play key roles in peripheral tolerance and prevention of autoimmune disease in mice as PD-1 knockout animals show dilated cardiomyopathy, splenomegaly, and loss of peripheral tolerance. Induced PD-L1 expression is common in many tumors including squamous cell carcinoma, colon adenocarcinoma, and breast adenocarcinoma. PD-L1 overexpression results in increased resistance of tumor cells to CD8 T cell mediated lysis. In mouse models of melanoma, tumor growth can be transiently arrested via treatment with antibodies which block the interaction between PD-L1 and its receptor PD-1. For these reasons anti-PD-1 mediated immunotherapies are currently being explored as cancer treatments. RecombiMAb anti-mouse PD-1 (CD279) (D265A) Specifications IsotypeMouse IgG1, κ Recommended Isotype Control(s)RecombiMAb mouse IgG1 (D265A) isotype control, anti-hen egg lysozyme Recommended Dilution BufferInVivoPure pH 7.0 Dilution Buffer ImmunogenRecombinant PD-1-Ig fusion protein Reported Applicationsin vivo blocking of PD-1/PD-L signaling*in vitro PD-1 neutralization*Immunohistochemistry (frozen)*Immunofluorescence*Western blot*Flow cytometry**Reported for the original rat IgG2a 29F.1A12 antibody FormulationPBS, pH 7.0Contains no stabilizers or preservatives Endotoxin<1EU/mg (<0.001EU/μg) Determined by LAL gel clotting assay Aggregation<5% Determined by SEC Purity>95%Determined by SDS-PAGE Sterility0.2 μm filtration ProductionPurified from HEK293 cell supernatant in an animal-free facility PurificationProtein G Molecular Weight150 kDa StorageThe antibody solution should be stored at the stock concentration at 4°C. Do not freeze.