We use cookies to understand how you use our site and to improve your experience. This includes personalizing content and advertising. To learn more, click here. By continuing to use our site, you accept our use of cookies. Cookie Policy.

Features Partner Sites Information LinkXpress
Sign In
Advertise with Us
Technopath Clinical Diagnostics - An LGC Company

PERKIN ELMER CORPORATION

PerkinElmer provides detection technologies to detect and identify diseases, imaging technologies to help visualize s... read more Featured Products: More products

Download Mobile App




Events

ATTENTION: Due to the COVID-19 PANDEMIC, many events are being rescheduled for a later date, converted into virtual venues, or altogether cancelled. Please check with the event organizer or website prior to planning for any forthcoming event.

Simple Colorimetric Assay Determines Human Mercaptalbumin Concentration

By LabMedica International staff writers
Posted on 30 Jun 2022
Print article
Image: VICTOR Nivo Multimode Microplate Reader (Photo courtesy of PerkinElmer)
Image: VICTOR Nivo Multimode Microplate Reader (Photo courtesy of PerkinElmer)

Human serum albumin (HAS) can take on two forms, mercaptalbumin (HMA) or non-mercaptalbumin (HNA), depending on the redox status of its Cys34. The ratio of HMA and HNA is considered to be a novel biomarker of oxidative stress.

The majority of studies on the redox state of HSA utilize high performance liquid chromatography (HPLC). Several recent studies have also employed mass spectrometry, which yields a higher resolution, thus allowing for finer distinctions between molecular structures. A handful of studies have explored alternative approaches using simple dye-based assays.

Clinical Laboratorians at The University of Tokyo Hospital (Tokyo, Japan) collaborated with colleagues at Nitto Boseki Co. Ltd., (Kawasaki, Japan) measured the concentrations of HMA in serum samples from 36 healthy subjects based on absorption changes of Michler's Hydrol. Michler's Hydrol (4,4′-Bis(dimethylamino)benzhydrol) is a blue dye with a maximum absorption at 612 nm, and its absorption decreases when it reacts with a thiol group.

After processing the samples, the solution was incubated further at room temperature for 20 minutes and absorbance at 612 nm (A612) was measured by a 96-well plate Nivo 3F microplate reader (PerkinElmer, Waltham, MA, USA). To evaluate the performance of the assays, the correlation was determined by a linear regression between the HMA concentrations obtained by Michler's Hydrol assay and corresponding reference concentrations obtained by HPLC and Bromocresol purple (BCP) assays.

The scientists reported that HMA concentrations obtained by Michler's Hydrol assay were highly correlated (r2 = 0.97) with reference values obtained by HPLC (HMA %) and BCP assay (total albumin). The HNA% obtained by Michler's Hydrol and BCP assays combined also gave a good correlation (r2 = 0.96) and a small deviation (average 2.4%) with respect to HPLC as a reference method. The use of the HMA/HNA ratio, which is a function of oxidation of Cys34, is considered a potential biomarker to assess the severity of chronic diseases for which oxidative stress is a significant etiological manifestation.

The authors concluded that the concentrations of HMA in human serum can be measured by Michler's Hydrol assay, which is based on an approximation of HMA concentrations with Michler's Hydrol's colorimetric changes mainly due to thiols. The assay accuracy was significantly improved by pre-incubation of a subset of the samples with an alkylating agent (iodoacetamide) and subtracting their absorption from the absorption of corresponding samples without alkylation. Proportions of HMA and HNA were also obtained by combining Michler's Hydrol assay and the BCP assay (for total albumin concentration). The results correlated well with HPLC, which has been established as a standard method for HMA/HNA measurement. The study was published on June 8, 2022 in the journal Practical Laboratory Medicine.

Related Links:
The University of Tokyo Hospital
Nitto Boseki Co. Ltd
PerkinElmer

 

Gold Supplier
Real-Time PCR System
Applied Biosystems QuantStudio 7 Pro Dx
New
Automated Nucleic Acid Extraction Instrument
DA3500
New
Microtome
M-250
New
Lab Incubator
IN-601

Print article
SUGENTECH INC.

Channels

Clinical Chem.

view channel
Image: ELISA kit for liver-type fatty acid–binding protein (L-FABP). The level of L-FABP present in urine reflects the level of renal tubular dysfunction (Photo courtesy of Sekisui Medical Co)

Urinary Biomarkers Predict Weaning From Acute Dialysis Therapy

Acute kidney injury is associated with a higher risk of chronic kidney disease (CKD), end-stage renal disease, and long-term adverse cardiovascular effects. Critically ill patients with acute kidney injury... Read more

Microbiology

view channel
Image: Ring-form trophozoites of Plasmodium vivax in a thin blood smear (Photo courtesy of Centers for Disease Control and Prevention)

Immune Regulators Predict Severity of Plasmodium vivax Malaria

Cytokines and chemokines are immune response molecules that display diverse functions, such as inflammation and immune regulation. In Plasmodium vivax infections, the uncontrolled production of these molecules... Read more

Pathology

view channel
Image: Breast cancer spread uncovered by new molecular microscopy (Photo courtesy of Wellcome Sanger Institute)

New Molecular Microscopy Tool Uncovers Breast Cancer Spread

Breast cancer commonly starts when cells start to grow uncontrollably, often due to mutations in the cells. Overtime the tumor becomes a patchwork of cells, called cancer clones, each with different mutations.... Read more

Industry

view channel
Image: With Cell IDx’s acquisition, Leica Biosystems will be moving its multiplexing menu forward (Photo courtesy of Leica Biosystems)

Leica Biosystems Acquires Cell IDx, Expanding Offerings in Multiplexed Tissue Profiling

Leica Biosystems, a technology leader in automated staining and brightfield and fluorescent imaging (Nussloch, Germany), has acquired Cell IDx, Inc. (San Diego, CA, USA), which provides multiplex staining... Read more
Copyright © 2000-2022 Globetech Media. All rights reserved.