Total protein is an effective loading control for cerebrospinal fluid western blots

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Total protein is an effective loading control for cerebrospinal fluid western blots

BACKGROUND Cerebrospinal fluid (CSF) has been used to identify biomarkers of neurological disease. CSF protein biomarkers identified by high-throughput methods, however, require further validation. While Western blotting (WB) is well-suited to this task, the lack of a validated loading control for CSF WB limits the method's accuracy. NEW METHOD We investigated the use of total protein (TP) as...

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Estimation of reference intervals for total protein in cerebrospinal fluid.

Protein in cerebrospinal fluid (CSF) was measured by a modified biuret procedure and in two automated instruments, the Du Pont aca and the Kodak Ektachem. The latter's dry-slide reagent was also evaluated for precision, linearity, and effect of potential interferents. In vitro, ampicillin and vancomycin increase the apparent value for CSF protein as measured with the Ektachem slides. We exclude...

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Total Protein Analysis as a Reliable Loading Control for Quantitative Fluorescent Western Blotting

Western blotting has been a key technique for determining the relative expression of proteins within complex biological samples since the first publications in 1979. Recent developments in sensitive fluorescent labels, with truly quantifiable linear ranges and greater limits of detection, have allowed biologists to probe tissue specific pathways and processes with higher resolution than ever be...

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Is standardization more important than methodology for assay of total protein in cerebrospinal fluid?

Four manual micromethods for protein determination, two turbidimetric (trichloroacetic and sulfosalicylic acid-sodium sulfate) and two colorimetric (Lowry and Coomassie Brilliant Blue--sodium dodecyl sulfate, CBB-SDS) were used to compare the standard curves for total protein (0.30 to 3 g/L) produced with three reference materials: bovine serum albumin, human serum albumin, and diluted human se...

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ژورنال

عنوان ژورنال: Journal of Neuroscience Methods

سال: 2015

ISSN: 0165-0270

DOI: 10.1016/j.jneumeth.2015.05.011