FKBP51 was immunoprecipitated using 0.5mg HL60 whole cell extract, 5µg of Rabbit polyclonal to FKBP51 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, HL60 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab46002. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697). Band: 52kDa: FKBP51.
IHC image of FKBP51 staining in human breast carcinoma FFPE section, performed on a BondTM 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 ab46002, 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.
Immunocytochemistry/ Immunofluorescence - FKBP51 antibody (ab46002)This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.
ab46002 staining human U2OS, mouse MEF, and rat RIE1 cells by ICC/IF. Cells were PFA fixed and permeabilized in 0.2% Triton-100 [PBS] prior to blocking in 1% BSA for 15 minutes at 20°C. The primary antibody was diluted 1/100 and incubated with the samples for 45 minutes at 20°C. The secondary antibody was an Alexa Fluor® 647 conjugated donkey anti-rabbit, diluted 1/1000.
Rotoli D et al. Commitment of Scaffold Proteins in the Onco-Biology of Human Colorectal Cancer and Liver Metastases after Oxaliplatin-Based Chemotherapy. Int J Mol Sci18:N/A (2017).
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Rotoli D et al. Expression and localization of the immunophilin FKBP51 in colorectal carcinomas and primary metastases, and alterations following oxaliplatin-based chemotherapy. Oncol Lett12:1315-1322 (2016).
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