JAK2 Monoclonal Antibody (691R5)

Protocol tips

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	-Mouse (assay dependent)

Protocol tips
-Mouse (assay dependent)

Publication protocol

Purified Jak2 and fos-GHRct-fos were incubated in 50mM HEPES (pH 7.5), 0.5mM dithiothreitol, 0.1M NaCl, and 5mM MnCl2 at 37°C in a total volume of 60 μL. After 20 minutes, 10-μL aliquots were sampled for input analysis, 2-μL anti-GHR serum was added, and the incubation was continued on ice. After 2 hours, protein A beads were added, and after 1 hour on ice, the beads were washed twice with 400-μL PBS containing 2% BSA and 0.5% Tween 20, and once with 500-μL PBS. The beads were boiled in sample buffer, and the proteins were separated on SDS-PAGE gels and analyzed by Western blotting using anti-GHR and anti-Jak2 (mouse). Detection and quantification of the signals were performed with an Odyssey system (LI-COR Biosciences).

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Manufacturer protocol

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