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Synthetic peptide conjugated to KLH derived from within residues 500 to the C-terminus of Human Egr1.
(Peptide available as ab56196.)
Our Abpromise guarantee covers the use of ab55160 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 5 µg/ml. Detects a band of approximately 55, 85 kDa (predicted molecular weight: 58 kDa).Can be blocked with Human Egr1 peptide (ab56196).|
|IP||Use a concentration of 5 µg/ml.|
|IHC-P||1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|Flow Cyt||Use 1µg for 106 cells.|
Egr1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Egr1 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 ab55160.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 55kDa; Egr1
ab55160 (1/250) staining Egr1 in paraffin-embedded Human placenta tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. The secondary antibody was goat anti rabbit/mouse conjugated to HRP. For further experimental details please refer to abreview.
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