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Our Abpromise guarantee covers the use of ab53034 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||1/500 - 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).|
|IHC-P||Use a concentration of 2 µg/ml.|
p15 INK4b was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to p15 INK4b 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, Hela 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 ab53034.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 15kDa; p15 INK4b
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