Eblen, Scott T. ; Vinay Kumar, N. ; Weber, Michael J. (2007) Using genetically engineered kinases to screen for novel protein kinase substrates: phosphorylation of substrates in cell lysates with exogenous kinase Cold Spring Harbor Protocols, 2007 (8). No pp. given. ISSN 1559-6095
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Official URL: http://cshprotocols.cshlp.org/content/2007/4/pdb.p...
Related URL: http://dx.doi.org/10.1101/pdb.prot4639
Abstract
This protocol describes a method for detection of direct substrates of a protein kinase in cell lysates or fractions. The approach involves the addition of recombinant mutant kinase and [γ-32P]ATP analog to cell lysates. This technique has been successfully used for the phosphorylation of Extracellular signal-regulated Kinase 2 (ERK2) substrates in cell lysates from SKOV-3 ovarian cells; however, the methodology can be applied to other protein kinases as well.
| Item Type: | Article |
|---|---|
| Source: | Copyright of this article belongs to Cold Spring Harbor Laboratory Press. |
| ID Code: | 113518 |
| Deposited On: | 25 May 2018 07:21 |
| Last Modified: | 25 May 2018 07:21 |
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