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A synthetic peptide corresponding to 14 amino acids near the carboxy terminus of human IRF7
Our Abpromise guarantee covers the use of ab62505 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 51 kDa (predicted molecular weight: 54 kDa).Can be blocked with Human IRF7 peptide (ab92594).|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 5 µg/ml.|
ICC/IF image of ab62505 stained HepG2 cells (ab7900). The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab62505, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IRF7 was immunoprecipitated using 0.5mg Jurkat whole cell extract (ab7899), 5µg of Rabbit polyclonal to IRF7 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, Jurkat 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab62505.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 49kDa: IRF7; non specific - 40kDa: We are unsure as to the identity of this extra band.