Our Abpromise guarantee covers the use of ab59542 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/4000. Predicted molecular weight: 31 kDa.|
|ELISA||1/1000 - 1/4000.|
|IHC-Fr||1/500 - 1/1000.|
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||1/50 - 1/100.|
|IP||Use at an assay dependent concentration.|
Peroxiredoxin 4 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µl of Rabbit polyclonal to Peroxiredoxin 4 sera 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, Hek293 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 ab59542.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 27kDa; Peroxiredoxin 4, non specific - as present in control (lane 2); 27kDa: We are confident this was due to slight lane contamination and the band seen in the IP lane is our target of interest.