Product Information
Cells are lysed in a urea-containing buffer, cellular proteins are digested by proteases, and the resulting peptides are purified by reversed-phase, solid-phase extraction. Peptides are then subjected to immunoaffinity purification using a PTMScan® antibody conjugated to protein A agarose beads. Unbound peptides are removed through washing, and the captured PTM-containing peptides are eluted with dilute acid. Reversed-phase purification is performed on microtips to desalt and separate peptides from antibody prior to concentrating the enriched peptides for LC-MS/MS analysis. CST recommends the use of PTMScan® IAP Buffer #9993 included in the kit. An alternate PTMScan® IAP Buffer Plus Detergent #9992 which may reduce nonspecific interactions is available separately. A detailed protocol and Limited Use License allowing the use of the patented PTMScan® method is included with the kit.
For more information on our PTMScan® Motif Antibody Kits, please visit www.cellsignal.com/support. Background information specific to each component in the sampler kit can be found on the individual webpages. Phospho-PKA Substrate Motif is described on the webpage for PTMScan® Phospho-PKA Substrate Motif (RRXS*/T*) Kit #5565. Phospho-AMPK Substrate Motif is described on the webpage for PTMScan® Phospho-AMPK Substrate Motif (LXRXXS*/T*) Kit #5564. Phospho-Akt Substrate Motif is described on the webpage for PTMScan® Phospho-Akt Substrate Motif mAb 1 (RXXS*/T*) Kit #5561. Phospho-CK2 Substrate Motif is described on the webpage for PTMScan® Phospho-CK2 Substrate Motif (S*/T*DXE) Kit #12170. Phospho-Tyrosine is described on the webpage for PTMScan® Phospho-Tyrosine Rabbit mAb (P-Tyr-1000) Kit #8803.
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