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AEEQKLISEEDLconjugated to KLH, corresponding to amino acids 408-420 of Human c-Myc.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Our Abpromise guarantee covers the use of ab56 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent concentration. 2ul per 500ug of extract, see H. Zhang et al.|
|WB||1/500 - 1/1000. Predicted molecular weight: 49 kDa. Additional non-specific bands observed at 75, 110, 140 kDa using mouse and human cells (see Abreview).|
|IP||Use a concentration of 5 µg/ml.|
|IHC-P||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
c-Myc was immunoprecipitated using 0.5mg CHO overexpressing Stra8 whole cell lysate, 5µg of Mouse monoclonal to c-Myc 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, CHO overexpressing Stra8 whole cell lysate 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 ab56.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.
Band: 49KDa; c-Myc
Overlay histogram showing HL60 cells stained with ab56 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Image courtesy of an anonymous Abreview.