The kit offers a set of six different MAbs specific for serine phosphorylation sites.
Phosphorylation patterns in a given cell extract may differ when probed with different antibodies due to sequence specificity. Recognition therefore depends on two criteria: phosphorylation and surrounding aa motif. If one of the two criteria is not fulfilled, the antibody will not detect the phosphorylation site. Specificity has been determined by epitope mapping using degenerated phosphopeptide libraries. Clone 16B4 strongly interacts with pSK and pSP motifs, thus specifically recognizes substrates of MAP/SAP Kinases and CDC kinases.Clones 1C8, 4A3, 4A9, 4H4 and 7F12 interact with epitopes containing phosphoserine in the context of positive or neutral aa (e.g. substrates of PKA, PKB, PKC, PKG etc.) Though revealing a similar pattern in epitope mapping using phosphopeptide libraries, these antibodies interact with different sets of phosphoproteins in whole cell lysates.
Immunoprecipitation, Western blot (20µl/lane (mini gel) for HRPO/DAB detection, or 5µl/lane (mini gel) for HRPO/ECL detection).
Do not use milk/casein for blocking and dilution! Replace by BSA! We recommend to use 1% BSA, 1% PVP-10 (polyvinyl-pyrrolidone), 1% PEG 3500, 0.2% Tween 20 in 2x PBS.
Optimal conditions must be determined individually for each application.
Ordering information: Contents: Contains 25µg each of the following MAbs:
Clone 1C8 (Prod. No. ALX-804-162)
Clone 4A3 (Prod. No. ALX-804-163)
Clone 4A9 (Prod. No. ALX-804-164)
Clone 4H4 (Prod. No. ALX-804-165)
Clone 7F12 (Prod. No. ALX-804-166)
Clone 16B4 (Prod. No. ALX-804-167)