HEK293T cells cultured in 10 cm dish were transfected with 5 μg Flag-RECQL5 using PEI (Polysciences Inc). After 48 h, harvest the cells and lysate them in NETN lysis buffer. The Flag M2 beads (Sigma) were washed with NETN lysis buffer. The protein A/G beads were washed and then mixed with Rabbit IgG as negative control or USP28 antibodies. About 2 mg of total protein from the cell lysates were incubated with the beads prepared above for 3 h at 4 °C. The bound proteins were denatured by adding 50 μl SDS loading buffer and boiling for 5 min and then analyzed by immunoblotting.
Flag recql5
Manufactured by Polysciences
Sourced in United States
Flag-RECQL5 is a laboratory reagent that contains a recombinant human RECQL5 protein tagged with a FLAG epitope. RECQL5 is a DNA helicase that plays a role in DNA repair and maintenance. This product is intended for use in research applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using flag recql5
1
Immunoprecipitation of RECQL5 and USP28
2
Flag-RECQL5 Immunoprecipitation in HEK293T
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HEK293T cells cultured in 10 cm dish were transfected with 5 μg Flag-RECQL5 using PEI (Polysciences Inc, Warrington, PA, USA). After 48hrs, harvest the cells and lysate them in NETN lysis buffer. The Flag M2 beads (Sigma) were washed with NETN lysis buffer. The protein A/G beads were washed and then mixed with Rabbit IgG as negative control or USP28 antibodies. About 2 μg total protein from the cell lysates were incubated with the beads prepared above for 3 hours at 4℃. The bound proteins were denatured by adding 50 μl SDS loading buffer and boiling for 5 minutes, and then analyzed by immunoblotting.
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