Replication protein A purified from U293 cells.
Our Abpromise guarantee covers the use of ab16850 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC||Use a concentration of 1 µg/ml.|
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|IP||Use a concentration of 5 µg/ml.|
|ICC/IF||Use at an assay dependent concentration.|
RPA32/RPA2 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Mouse monoclonal to RPA32/RPA2 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, HepG2 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 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab16850.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.
Band: 29kDa; RPA32/RPA2
IHC image of RPA32/RPA2 staining in Human normal lung 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 ab16850, 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.
Staining of SKN cells with ab16850 (red) at a dilution of 1/1000. The cells were fixed in paraformaldehyde for 10 minutes prior to incubation with ab16850. The DNA is stained with DAPI (blue). 100x magnification.