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Our Abpromise guarantee covers the use of ab58918 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use a concentration of 5 µg/ml.|
|ELISA||Use at an assay dependent concentration.
Titre using peptide based assay, 1:312500.
|WB||Use a concentration of 5 - 10 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 37 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.|
Connexin 37 / GJA4 was immunoprecipitated using 0.5mg NIH3T3 whole cell extract, 5µg of Rabbit polyclonal to Connexin 37 / GJA4 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, NIH3T3 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 ab58918.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 45kDa; Connexin 37 / GJA4
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