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  • Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS).

Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS).

Metabolites (2012-01-01)
Mathias Haag, Angelika Schmidt, Timo Sachsenheimer, Britta Brügger
ABSTRACT

Lipids, such as phosphoinositides (PIPs) and diacylglycerol (DAG), are important signaling intermediates involved in cellular processes such as T cell receptor (TCR)-mediated signal transduction. Here we report identification and quantification of PIP, PIP2 and DAG from crude lipid extracts. Capitalizing on the different extraction properties of PIPs and DAGs allowed us to efficiently recover both lipid classes from one sample. Rapid analysis of endogenous signaling molecules was performed by nano-electrospray ionization tandem mass spectrometry (nano-ESI MS/MS), employing lipid class-specific neutral loss and multiple precursor ion scanning for their identification and quantification. Profiling of DAG, PIP and PIP2 molecular species in primary human T cells before and after TCR stimulation resulted in a two-fold increase in DAG levels with a shift towards 1-stearoyl-2-arachidonoyl-DAG in stimulated cells. PIP2 levels were slightly reduced, while PIP levels remained unchanged.

MATERIALS
Product Number
Brand
Product Description

Avanti
Brain PI(4,5)P2, Avanti Research - A Croda Brand
Avanti
18:1 DG, Avanti Research - A Croda Brand
Avanti
16:0-18:1 DG, 1-palmitoyl-2-oleoyl-sn-glycerol, chloroform
Avanti
18:0-20:4 DG, 1-stearoyl-2-arachidonoyl-sn-glycerol, chloroform
Avanti
16:0-18:1 DG, 1-palmitoyl-2-oleoyl-sn-glycerol, neat oil
Avanti
16:0 DG, 1,2-dipalmitoyl-sn-glycerol, chloroform
Avanti
16:0 DG, 1,2-dipalmitoyl-sn-glycerol, powder
Avanti
18:1 DG, 1-2-dioleoyl-sn-glycerol, chloroform
Avanti
Brain PI(4,5)P2, Avanti Research - A Croda Brand
Avanti
17:0-20:4 PI(4)P, Avanti Research - A Croda Brand LM1901, powder
Avanti
20:0 PC, 1,2-diarachidoyl-sn-glycero-3-phosphocholine, powder
Avanti
20:0 PC, 1,2-diarachidoyl-sn-glycero-3-phosphocholine, chloroform
Avanti
21:0 PC, 1,2-diheneicosanoyl-sn-glycero-3-phosphocholine, powder
Avanti
Brain PI(4)P, Avanti Research - A Croda Brand 840045P, powder
Avanti
Brain PI(4)P, Avanti Research - A Croda Brand 840045X
Avanti
14:0 PC (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine, chloroform
Avanti
13:0 PC, 1,2-ditridecanoyl-sn-glycero-3-phosphocholine, chloroform
Avanti
14:0 PC (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine, powder
Avanti
14:0 DG, 1,2-dimyristoyl-sn-glycerol, powder
Avanti
13:0 PC, 1,2-ditridecanoyl-sn-glycero-3-phosphocholine, powder