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Programmed cell death protein 1, also known as PD-1 and CD279 (cluster of differentiation 279) or PDCD1, is a protein that in humans is encoded by the PDCD1 gene. Mature Cynomolgus monkey PD-1 consists of a 148 amino acid (aa) extracellular region (ECD) with one immunoglobulin-like V-type domain, a 24 aa transmembrane domain, and a 95 aa cytoplasmic region. The Cynomolgus monkey PD-1 ECD shares 95% aa sequence identity with the human PD-1 ECD. The cytoplasmic tail contains two tyrosine residues that form the immunoreceptor tyrosine-based inhibitory motif (ITIM) and immunoreceptor tyrosine-based switch motif (ITSM) that are important for mediating PD-1 signaling. PD‑1 is expressed on activated T cells, B cells, monocytes, and dendritic cells while. PD-L1 expression is constitutive on the same cells and also on nonhematopoietic cells such as lung endothelial cells and hepatocytes. Ligation of PD-L1 with PD-1 induces co-inhibitory signals on T cells promoting their apoptosis, anergy, and functional exhaustion.
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文献和实验Purification of Resistance Protein Complexes Using a Biotinylated Affinity (HPB) Tag
function. Many R proteins are associated with the plasma membrane (PM) and expressed at low levels. Here, we describe a method to purify such low-abundance PM R protein �complexes from Arabidopsis using a biotinylated affinity tag, called the HPB tag
Western Blotting with Biotinylated Antibodies
. 3.Centrifuge the homogenate (16,000 x g, 15 C) for 5 minutes to pellet insoluble material, then discard pellet. 4.Dilute an aliquot of the tissue lysate sample at least 10-fold for the BCA (Pierce) protein concentration assay. POLYACRYLAMIDE GEL
on the beads was detected using biotinylated antigen/streptavidin-phycoerythrin. The method was demonstrated by isotyping antibodies directed against an anti-CD11a antibody therapeutic (RAPTIVA� /efalizumab) from the serum of a cynomolgus monkey hyper-immunized
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