Our Abpromise guarantee covers the use of ab47010 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 37 kDa).|
|IP||Use a concentration of 5 µg/ml.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
Lane 1 :40 kDa
Lane 2:37 kDa
Lane 3:62 kDa
Lane 4: 37 kDa
LDHA was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to LDHA 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 ab47010.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 36kDa; LDHA
This image is courtesy of an anonymous Abreview
Blocking step: 5% Milk for 1 hour at 24ºC
Primary antibody incubation for 12 hours at 4ºC