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Tissue, cells or virus corresponding to Human PD1. YT cells (human T/NK cell Leukemia)
Database link: Q15116
Alternative versions available:
Our Abpromise guarantee covers the use of ab52587 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/50. Predicted molecular weight: 32 kDa.|
|Flow Cyt||Use at an assay dependent concentration.
|IHC-Fr||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 5 - 10 µg/ml.|
|IP||Use at an assay dependent concentration.|
|IHC-P||1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
ab52587 stained in MOLT4 cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab52587 at 10µg/ml and ab6046 (Rabbit polyclonal to beta Tubulin - Loading Control) at 1ug/ml overnight at +4°C. The secondary antibodies used were ab150177 used at 1 ug/ml (colored green) and ab150080 (pseudo-colored red) used at 2ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.
IHC image of PD1staining in normal human tonsil formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab52587, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Immunohistochemical analysis of frozen Human liver tissue labeling PD1 with ab52587 at 1/50 dilution.
This image is courtesy of an anonymous collaborator.