Two optimized combination assays to examine apoptosis pathways in clinical samples.

Two optimized combination assays to examine apoptosis pathways in clinical samples. Research Abstract Details 

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Two optimized combination assays to examine apoptosis pathways in clinical samples. Abstract Text:

Mark Hollier,Toni Whistler,Carolyn Dawson,Suzanne D Vernon,

BACKGROUND: A consequence of a number of diseases is an alteration in apoptosis. Currently, there is no single assay that measures the main stages of apoptosis, requiring that multiple assays be performed. This hinders studies on clinical samples that have limited cell numbers. Our objective was to combine and optimize assays that target specific stages of apoptosis for use in a typical clinical blood sample. METHODS: Two flow cytometric assays were developed for use on peripheral blood mononuclear cells (PBMC) collected in two 8-ml tubes from a single draw. One measures caspase-12 activity, the level of active caspase-3 and DNA fragmentation. The second assesses depolarization of the mitochondria and phosphatidylserine externalization. Cell populations present within the samples were determined by flow cytometry. Apoptosis was validated by ELISA. RESULTS: Each assay was optimized for use with cell numbers and sample volumes typical of clinical blood samples. Each combination assay effectively distinguished apoptotic from nonapoptotic blood cells. CONCLUSIONS: This combined optimized method comprised of two independent assays makes it possible to assay the major pathways of apoptosis in addition to determining the blood cell subsets that are affected.

Two optimized combination assays to examine apoptosis pathways in clinical samples. Publishing Authors By Initials

M Hollier,T Whistler,C Dawson,SD Vernon,

For similar phosphatidic acids: glycerophospholipids: phosphatidylserines research abstracts see: phosphatidic acids: glycerophospholipids: phosphatidylserines research

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Two optimized combination assays to examine apoptosis pathways in clinical samples. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Cytometry. Part A : the journal of the Internation

VOLUME: 71

Page Numbers: 675-85

Journal Abbreviation: Cytometry A

ISSN: 1552-4922

DAY: 28

MONTH: Sep

YEAR: 2007

Two optimized combination assays to examine apoptosis pathways in clinical samples. Information

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LANGUAGE: eng

NlmUniqueID: 101235694

Two optimized combination assays to examine apoptosis pathways in clinical samples. Keywords Mesh Terms:

KEYWORDS: Phosphatidylserines

MESH TERMS: metabolism

Chemical & Substance for Abstract: Two optimized combination assays to examine apoptosis pathways in clinical samples. Information

Substance Name: Caspase 3

Registry Number: EC 3.4.22.-

Grant and Affiliation Information for Two optimized combination assays to examine apoptosis pathways in clinical samples.

AFFILIATION: Chronic Viral Diseases Branch, Division of Viral and Rickettsial Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30329, USA.

Country: United States

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MEDLINETA: Cytometry A

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