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Application Notes
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When peptides are reconstituted in 200 μL water, the concentration would be also 0.5 mg/mL. To start, the best ratio would be 1:1 (which means molar excess of peptides relative to antibodies when identical volumes are mixed). Please mix equal volumes of peptide and antibody at the required dilution and leave at ambient temperature. Best is to have two identical blots, to be incubated with equal amount of antibodies, but one with the antibodies pre-adsorbed to the peptide for 20 min. Then incubate and develop in parallel the two blots.Approx 57 kDa band observed in lysates of cell lines HeLa, HepG2, and K562 (calculated MW of 57.3 kDa according to NP_002121.4). Recommended concentration: 1-3 μg/mL.
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