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Anti nek1 h3t1

Manufactured by Merck Group
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Anti-NEK1 (H3T1) is a laboratory reagent produced by Merck Group. It is an antibody that specifically targets the NEK1 (Never in Mitosis Gene A-Related Kinase 1) protein and the H3T1 (Histone H3 Threonine 1) epitope. This product is designed for use in various research applications, such as immunoassays and immunoblotting, to study the role of NEK1 and H3T1 in cellular processes.

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Lab products found in correlation

Microspin column, by GE Healthcare (2 mentions) Protein a g plus agarose bead, by Santa Cruz Biotechnology (2 mentions) Anti flag, by Merck Group (2 mentions) Complete protease inhibitor cocktail, by Roche (2 mentions) Complete protease inhibitor, by Roche (2 mentions)

2 protocols using anti nek1 h3t1

1

Affinity Purification of NEK1 Protein Complexes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein A/G PLUS-agarose beads (Santa Cruz Biotechnology) were washed three times in extraction buffer (10mM HEPES [pH7.9], 10mM NaCl, 3mM MgCl2, freshly added 1mM DTT, 1mM Na3VO4) supplemented with complete protease inhibitor cocktail (Roche) and then coupled to polyclonal anti-NEK1 (H3T1) or polyclonal anti-FLAG (Sigma, F7425) overnight at 4°C. Coupled beads were washed three times in extraction buffer and incubated with fresh bovine retinal lysates overnight at 4°C. Retinas were homogenized by sonication on ice two times for 30 seconds in extraction buffer (10mM HEPES [pH7.9], 10mM NaCl, 3mM MgCl2, freshly added 1mM DTT, 1mM Na3VO4) supplemented with complete protease inhibitor cocktail (Roche). Beads were washed five times with extraction buffer, transferred to microspin columns (GE Healthcare) and washed three times in washing buffer (1xTBS, 0.12% NP-40, supplemented with complete protease inhibitors (Roche)). Then, neutralization buffer (1M Tris/HCl, [pH8.0]) was added. Eluates were retrieved by adding 3x elution buffer (200 mM glycine [pH2.5]) for 20 min. at 4°C and subjected to protein precipitation with chloroform and methanol. Protein precipitates were subsequently subjected to mass spectrometry analysis and peptide identification as previously described48 (link).
Wheway G., Schmidts M., Mans D.A., Szymanska K., Nguyen T.M., Racher H., Phelps I.G., Toedt G., Kennedy J., Wunderlich K.A., Sorusch N., Abdelhamed Z.A., Natarajan S., Herridge W., van Reeuwijk J., Horn N., Boldt K., Parry D.A., Letteboer S.J., Roosing S., Adams M., Bell S.M., Bond J., Higgins J., Morrison E.E., Tomlinson D.C., Slaats G.G., van Dam T.J., Huang L., Kessler K., Giessl A., Logan C.V., Boyle E.A., Shendure J., Anazi S., Aldahmesh M., Al Hazzaa S., Hegele R.A., Ober C., Frosk P., Mhanni A.A., Chodirker B.N., Chudley A.E., Lamont R., Bernier F.P., Beaulieu C.L., Gordon P., Pon R.T., Donahue C., Barkovich A.J., Wolf L., Toomes C., Thiel C.T., Boycott K.M., McKibbin M., Inglehearn C.F., Stewart F., Omran H., Huynen M.A., Sergouniotis P.I., Alkuraya F.S., Parboosingh J.S., Innes A.M., Willoughby C.E., Giles R.H., Webster A.R., Ueffing M., Blacque O., Gleeson J.G., Wolfrum U., Beales P.L., Gibson T., Doherty D., Mitchison H.M., Roepman R, & Johnson C.A. (2015). An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes. Nature cell biology, 17(8), 1074-1087.
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2

Affinity Purification of NEK1 Protein Complexes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein A/G PLUS-agarose beads (Santa Cruz Biotechnology) were washed three times in extraction buffer (10mM HEPES [pH7.9], 10mM NaCl, 3mM MgCl2, freshly added 1mM DTT, 1mM Na3VO4) supplemented with complete protease inhibitor cocktail (Roche) and then coupled to polyclonal anti-NEK1 (H3T1) or polyclonal anti-FLAG (Sigma, F7425) overnight at 4°C. Coupled beads were washed three times in extraction buffer and incubated with fresh bovine retinal lysates overnight at 4°C. Retinas were homogenized by sonication on ice two times for 30 seconds in extraction buffer (10mM HEPES [pH7.9], 10mM NaCl, 3mM MgCl2, freshly added 1mM DTT, 1mM Na3VO4) supplemented with complete protease inhibitor cocktail (Roche). Beads were washed five times with extraction buffer, transferred to microspin columns (GE Healthcare) and washed three times in washing buffer (1xTBS, 0.12% NP-40, supplemented with complete protease inhibitors (Roche)). Then, neutralization buffer (1M Tris/HCl, [pH8.0]) was added. Eluates were retrieved by adding 3x elution buffer (200 mM glycine [pH2.5]) for 20 min. at 4°C and subjected to protein precipitation with chloroform and methanol. Protein precipitates were subsequently subjected to mass spectrometry analysis and peptide identification as previously described48 (link).
Wheway G., Schmidts M., Mans D.A., Szymanska K., Nguyen T.M., Racher H., Phelps I.G., Toedt G., Kennedy J., Wunderlich K.A., Sorusch N., Abdelhamed Z.A., Natarajan S., Herridge W., van Reeuwijk J., Horn N., Boldt K., Parry D.A., Letteboer S.J., Roosing S., Adams M., Bell S.M., Bond J., Higgins J., Morrison E.E., Tomlinson D.C., Slaats G.G., van Dam T.J., Huang L., Kessler K., Giessl A., Logan C.V., Boyle E.A., Shendure J., Anazi S., Aldahmesh M., Al Hazzaa S., Hegele R.A., Ober C., Frosk P., Mhanni A.A., Chodirker B.N., Chudley A.E., Lamont R., Bernier F.P., Beaulieu C.L., Gordon P., Pon R.T., Donahue C., Barkovich A.J., Wolf L., Toomes C., Thiel C.T., Boycott K.M., McKibbin M., Inglehearn C.F., Stewart F., Omran H., Huynen M.A., Sergouniotis P.I., Alkuraya F.S., Parboosingh J.S., Innes A.M., Willoughby C.E., Giles R.H., Webster A.R., Ueffing M., Blacque O., Gleeson J.G., Wolfrum U., Beales P.L., Gibson T., Doherty D., Mitchison H.M., Roepman R, & Johnson C.A. (2015). An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes. Nature cell biology, 17(8), 1074-1087.
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