Detection of TAPBPR and UGT1 in MHC Class I Complexes

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).

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Ilca F.T., Neerincx A., Hermann C., Marcu A., Stevanović S., Deane J.E, & Boyle L.H. (2018). TAPBPR mediates peptide dissociation from MHC class I using a leucine lever. eLife, 7, e40126.

Publication 2018

PeTe4 ab57411 H1029 mouse IgG2a isotype control

Amino acids Anti antibody Human Igg2a Isotype Mhc class i Monoclonal antibody Mouse Polyhistidine Rabbit Tapasin

Corresponding Organization :

Other organizations : University of Cambridge, University of Cape Town, University of Tübingen

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Variable analysis

independent variables

Use of PeTe4, a mouse monoclonal antibody (mAb) specific for the native conformation of TAPBPR, raised against amino acids 22–406 of human TAPBPR
Use of ab57411, a mouse mAb raised against amino acids 23–122 of TAPBPR that is reactive to denatured TAPBPR
Use of the rabbit mAb ab124879 to detect UGT1
Use of mAb HC10 to detect MHC class I heavy chains
Use of the mouse anti-polyhistidine mAb H1029 to detect soluble TAPBPR variants

dependent variables

Detection of TAPBPR
Detection of UGT1
Detection of MHC class I heavy chains
Detection of soluble TAPBPR variants

control variables

Use of a mouse IgG2a isotype control

controls

Positive control: PeTe4, a mouse monoclonal antibody (mAb) specific for the native conformation of TAPBPR
Negative control: Mouse IgG2a isotype control

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