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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.
|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.