Immunoaffinity-Based Liquid Chromatography Mass Spectrometric Assay to Accurately Quantify the Protein Concentration of HMGB1 in EDTA Plasma.

Szczegóły
Abstrakt

Tytuł:: Immunoaffinity-Based Liquid Chromatography Mass Spectrometric Assay to Accurately Quantify the Protein Concentration of HMGB1 in EDTA Plasma.
Autorzy:: Anselm V; Signatope GmbH, Reutlingen, Germany.
Steinhilber A; Signatope GmbH, Reutlingen, Germany.
Sommersdorf C; Signatope GmbH, Reutlingen, Germany.
Poetz O; Signatope GmbH, Reutlingen, Germany. .
Źródło:: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2261, pp. 277-289.
Typ publikacji:: Journal Article
Język:: English
Imprint Name(s):: Publication: Totowa, NJ : Humana Press
Original Publication: Clifton, N.J. : Humana Press,
MeSH Terms:: Analytic Sample Preparation Methods*
Blood Specimen Collection*
Chromatography, Affinity*
Proteomics*
Tandem Mass Spectrometry*
Edetic Acid/*pharmacology
HMGB1 Protein/*blood
Anticoagulants/pharmacology ; Humans ; Immunoprecipitation ; Proteolysis
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Contributed Indexing:: Keywords: Antibody; HMGB1; Immunoaffinity liquid chromatography mass spectrometric assay; Immunoprecipitation; Parallel reaction monitoring; Peptide quantification
Substance Nomenclature:: 0 (Anticoagulants)
0 (HMGB1 Protein)
0 (HMGB1 protein, human)
9G34HU7RV0 (Edetic Acid)
Entry Date(s):: Date Created: 20210109 Date Completed: 20210331 Latest Revision: 20210331
Update Code:: 20240105
DOI:: 10.1007/978-1-0716-1186-9_17
PMID:: 33420996
: Czasopismo naukowe

Targeted protein quantification can be challenging in body fluids such as plasma with regard to sensitivity and selectivity. In this chapter, we present a protocol for the quantification of high mobility group box 1 protein (HMGB1) in plasma using an immunoaffinity liquid chromatography mass spectrometric assay (IA-LC-MSMS). The protocol provides detailed assay instructions involving sample proteolysis, peptide-targeted immunoprecipitation, and LC-MSMS-based read out.

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