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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.
|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