有哪些信誉好的足球投注网站a high-sensitivity method for detection and measurement of hmgb1 protein concentration by high-affinity binding to dna hemicatenanes一个高灵敏度的检测和测量方法的高亲和性结合dna hemicatenanes hmgb1蛋白浓度.pdfVIP
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a high-sensitivity method for detection and measurement of hmgb1 protein concentration by high-affinity binding to dna hemicatenanes一个高灵敏度的检测和测量方法的高亲和性结合dna hemicatenanes hmgb1蛋白浓度
A High-Sensitivity Method for Detection and
Measurement of HMGB1 Protein Concentration by High-
Affinity Binding to DNA Hemicatenanes
´ ¨ ´
Claire Gaillard, Chloe Borde, Joel Gozlan, Vincent Marechal, Franc¸ois Strauss*
´ ´
Centre de Recherche des Cordeliers, Universite Pierre et Marie Curie, Universite Paris Descartes, INSERM, Paris, France
Abstract
Background: Protein HMGB1, an abundant nuclear non-histone protein that interacts with DNA and has an architectural
function in chromatin, was strikingly shown some years ago to also possess an extracellular function as an alarmin and a
mediator of inflammation. This extracellular function has since been actively studied, both from a fundamental point of view
and in relation to the involvement of HMGB1 in inflammatory diseases. A prerequisite for such studies is the ability to detect
HMGB1 in blood or other biological fluids and to accurately measure its concentration.
Methodology/Principal Findings: In addition to classical techniques (western blot, ELISA) that make use of specific anti-
HMGB1 antibodies, we present here a new, extremely sensitive technique that is based on the fact that hemicatenated DNA
loops (hcDNA) bind HMGB1 with extremely high affinity, higher than the affinity of specific antibodies, similar in that respect
to DNA aptamers. DNA-protein complexes formed between HMGB1 and radiolabeled hcDNA are analyzed by
electrophoresis on nondenaturing polyacrylamide gels using the band-shift assay method. In addition, using a simple
and fast protocol to purify HMGB1 on the basis of its solubility in perchloric acid allowed us to increase the sensitivity by
suppressing any nonspecific backgr
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