Metabolomic Analysis of Cell Cultures

We measured glucose and lactate concentrations enzymatically using the YSI 2500 Glucose/Lactate Analyzer (Yellow Springs, OH, USA). We measured glutamine concentration and enrichment by LC-MS/MS (AbSCIEX 6500 QTRAP with a Shimadzu ultrafast liquid chromatography system in negative ion mode) as we have previously reported [25 (link)] after spiking with [²H₄] taurine as an internal standard. We measured citrate, succinate, and malate concentration and enrichment using the same LC-MS/MS method. Mass/charge ratios for the unlabeled metabolites are shown in Table 2. Due to space constraints, the labeled intermediate mass/charge ratios are not shown.Table 2

Mass/charge ratios for the unlabeled metabolites

Analyte	Mass/charge
Citrate	191/173
Glutamate	146/128
Malate	133/115
Succinate	117/99

We measured amino acid concentrations with gas chromatography/mass spectrometry after derivatization as described previously [26 (link)]. We measured non-esterified fatty acid (NEFA) concentrations using the Wako NEFA-HR assay (Mountain View, CA, USA).

Free full text: Click here

Zhang X., Halberstam A.A., Zhu W., Leitner B.P., Thakral D., Bosenberg M.W, & Perry R.J. (2022). Isotope tracing reveals distinct substrate preference in murine melanoma subtypes with differing anti-tumor immunity. Cancer & Metabolism, 10, 21.

Publication 2022

AbSCIEX 6500 QTRAP NEFA-HR assay

Amino acid Assay Citrate Esterified fatty acid Gas chromatography mass spectrometry Glucose Glutamine Lactate Liquid chromatography Malate Ms ms Springs Succinate Taurine

Corresponding Organization :

Other organizations : Yale University

Top 5 similar protocols

Variable analysis

independent variables

None explicitly mentioned

dependent variables

Glucose concentration
Lactate concentration
Glutamine concentration and enrichment
Citrate, succinate, and malate concentration and enrichment
Amino acid concentrations
Non-esterified fatty acid (NEFA) concentrations

control variables

None explicitly mentioned

controls

Positive control: [2H4] taurine used as an internal standard for glutamine, citrate, succinate, and malate measurements by LC-MS/MS

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!