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Our Abpromise guarantee covers the use of ab65844 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 1 - 2 µg/ml. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 39 kDa.|
|IP||Use a concentration of 5 µg/ml.|
Annexin A1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Annexin A1 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 70C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab65844.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 39kDa; Annexin A1, non specific bands - 52kDa and in control (lane 2): We are unsure as to the identity of this extra band.
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