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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.|
|ICC/IF||Use at an assay dependent concentration.|
Immunofluorescence analysis of HeLa cells, using Anti-p15 INK4b antibody (ab53034). The picture on the right is blocked with the synthesized peptide.
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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