Enhanced detection and desalting free protocol for phosphopeptides eluted from immobilized Fe (III) affinity chromatography in direct MALDI TOF analysis

Triplin, a small molecule, reveals copper ion transport in ethylene signaling from ATX1 to RAN1

“…For these assays, 100 μl of the metal ions (100 μM or 10 mM) and 100 μl triplin (100 μM or10 mM) were mixed for MALDI-TOF-MS analysis according to [46]. A time-of–flight Axima Performance mass spectrometer (Shimadzu, Japan) was used.…”

Improving detection sensitivity of amino acids in thyroid tissues by using phthalic acid as a mobile phase additive in hydrophilic interaction chromatography-electrospray ionization-tandem mass spectrometry

“…Some of these carboxylic acids have been reported for the enhanced detection sensitivity of non-phosphorylated peptides by adding them to the sample solution or the MALDI matrix [37][38][39][40]. Thus utilization of these carboxylic acids as mobile phase additives was supposed to have the same effect on signal intensity of AAs [33,34].…”

Scalable, Non-denaturing Purification of Phosphoproteins Using Ga3+-IMAC: N2A and M1M2 Titin Components as Study case

“…IMAC systems have been reported that are based on Ga 3+ , Fe 3+ , Ti 4+ , Zn 2+ and Al 3+ ions (e.g. [8][9][10][11][12][13]) and include commercial products such as Fe 3+ -based PHOS-Select™ IMAC (Sigma) and, the now discontinued, Ga +3 -IMAC PhosphoProfile™ (Sigma). Traditionally, Fe 3+ has been the preferred immobilized ion for phosphor-group capture, but Ga 3+ has been shown to be superior in its binding specificity of phosphorylated species [8,9,13,14].…”

“…In fact, IMAC protocols (as well as MOAC) are commonly performed in microscale column formats and on proteolized peptide mixtures using strongly denaturing chemical elutants (e.g. acetonitrile, dihydroxybenzoic acid, ammonium hydroxide, trifluoracetic acid, or phosphoric acid at highly acidic pH values; [8,[10][11][12][15][16][17][18]) as this yields the best performance.…”