Pete4

TAPBPR was detected using either PeTe4, a mouse monoclonal antibody (mAb) specific for the native conformation of TAPBPR, raised against amino acids 22–406 of human TAPBPR (Boyle et al., 2013 (link)) that does not cross-react with tapasin (Hermann et al., 2013 (link)), or ab57411, a mouse mAb raised against amino acids 23–122 of TAPBPR that is reactive to denatured TAPBPR (Abcam, UK). Tapasin was detected using Pasta-1 (Dick et al., 2002 (link)), or with R.gp48N, a rabbit polyclonal antibody specific for tapasin (Sadasivan et al., 1996 (link)) (kind gifts from Peter Cresswell, Yale University School of Medicine). MHC I heavy chains were detected using mAb HC10 (Stam et al., 1986 (link)) and mAb HCA2 (Stam et al., 1990 (link)). β₂m was detected using a rabbit polyclonal antibody (Dako, UK). Cell surface MHC I molecules were detected using W6/32, a pan-MHC I mAb that recognizes a conformational epitope on the α2 domain of MHC I, in a manner dependent on presence of β₂m and peptide (Barnstable et al., 1978 (link)). Calnexin was detected via western blot analysis using the rabbit polyclonal ADI-SPA-860 (Enzo Life Sciences, UK). A mouse IgG2a isotype control was also used as a control (Sigma-Aldrich).

TAPBPR was detected using either PeTe4, a mouse monoclonal antibody (mAb) specific for the native conformation of TAPBPR, raised against amino acids 22–406 of human TAPBPR (Boyle et al., 2013 (link)) that does not cross-react with tapasin (Hermann et al., 2013 (link)), or ab57411, a mouse mAb raised against amino acids 23–122 of TAPBPR that is reactive to denatured TAPBPR (Abcam, UK). UGT1 was detected using the rabbit mAb ab124879 (Abcam). MHC class I heavy chains were detected using mAb HC10 (Stam et al., 1986 ). Soluble TAPBPR variants were detected using the mouse anti-polyhistidine mAb H1029 (Sigma-Aldrich). A mouse IgG2a isotype control was also used as a control (Sigma-Aldrich).