For each condition, 10 µl anti-FLAG M2 magnetic beads (Sigma-Aldrich, M8823-1 ml), pre-equilibrated in lysis buffer (see above), were added to 700 µl of lysates and incubated overnight at 4°C on a rotating wheel. Samples were washed three times with lysis buffer and proteins were eluted upon addition of 50 µl of 1× BOLT LDS (Novex no. B0007) with 100 mM DTT and boiling at 95°C for 10 min. A total of 10 µl of the immunoprecipitated samples as well as 10 µl of lysates (input) were analysed by immunoblots.
Bolt lds
Manufactured by Thermo Fisher Scientific
The BOLT LDS is a laboratory equipment product from Thermo Fisher Scientific. It is designed for liquid delivery and storage purposes. The core function of the BOLT LDS is to accurately and precisely transfer liquids in a controlled manner within a laboratory setting.
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Lab products found in correlation
2 protocols using bolt lds
1
Immunoprecipitation of FLAG-tagged Proteins
2
Immunoprecipitation and Immunoblotting Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For each condition, 10 µl anti-FLAG M2 magnetic beads (Sigma-Aldrich, M8823-1ML), pre-equilibrated in lysis buffer (see above), were added to 700 µl of lysates and incubated overnight at 4 °C on a rotating wheel. Samples were washed three times with lysis buffer and proteins were eluted upon addition of 50 µl of 1X BOLT LDS (Novex #B007) with 100 mM DTT and boiling at 95 °C for 10 minutes. 10 µl of the immunoprecipitated samples as well as 10 µl of lysates (input) were analysed by immunoblots.
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