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Synthetic peptide conjugated to KLH derived from within residues 750 - 850 of Human ZO1 tight junction protein.
(Peptide available as ab93087.)
Our Abpromise guarantee covers the use of ab61357 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 0.1 - 1 µg/ml. Detects a band of approximately 240 kDa (predicted molecular weight: 195 kDa).
Abcam recommends using BSA as the blocking agent.
|IP||Use a concentration of 5 µg/ml.|
ZO1 tight junction protein was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Mouse monoclonal to 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, Hek293 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 ab61357.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.
Band: 240kDa, non specific band - 90kDa: We are unsure as to the identity of this extra band; ZO1 tight junction protein
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"