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Our Abpromise guarantee covers the use of ab25085 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 28 kDa.|
|IHC-P||1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IHC-Fr||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 1 - 5 µg/ml.|
Calbindin was immunoprecipitated using 0.5mg Rat Brain tissue extract, 5µg of Rabbit polyclonal to Calbindin 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, Rat Brain tissue 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 ab25085.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).<
Band: 28kDa; Calbindin
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