Quantification of Soluble or Insoluble Fractions of Leishmania Parasite Proteins in Microvolume Applications: A Simplification to Standard Lowry Assay

Joint Authors

Siriwardana, Yamuna
Deepachandi, Bhagya
Karunaweera, Nadira D.
Weerasinghe, Sudath
Andrahennadi, Thisira Priyantha
Wickramarachchi, Nadeeja
Soysa, Preethi

Source

International Journal of Analytical Chemistry

Issue

Vol. 2020, Issue 2020 (31 Dec. 2020), pp.1-8, 8 p.

Publisher

Hindawi Publishing Corporation

Publication Date

2020-03-13

Country of Publication

Egypt

No. of Pages

8

Main Subjects

Chemistry
Science

Abstract EN

Protein quantification is often an essential step in any research field that involves proteins.

Although the standard Lowry assay and its modifications are most abundantly used in protein quantification, the existing methods are rigid or often demonstrate nonlinearity between protein concentration and color intensity.

A method for fast and accurate qualitative and/or quantitative determination of total soluble/insoluble proteins or micro-well plate immobilized proteins isolated from Leishmania parasites in microvolumes was described in the current study.

Improvements in cost-effective techniques are necessary to increase the research outputs in resource-limited settings.

This method is a modification to the established Lowry assay for protein quantification.

Concentrations of unknown samples were calculated using a standard curve prepared using a standard series of bovine serum albumin (BSA).

The optimized reagents were 2 N NaOH (sodium hydroxide), 2% Na2CO3 (sodium carbonate), 1% CuSO4 (copper sulfate), 2% KNaC4H4O6 (potassium sodium tartrate), and 2 N Folin and Ciocalteu’s phenol.

This modified protein assay was sensitive for quantifying Leishmania proteins in a total crude extract or in a soluble fraction within the approximate range of 10–500 μg/ml (1–50 μg/assay) and showed a linearity between color intensity and concentration of the protein.

This is an easier, fast, and accurate method for quantifying proteins with microvolumes in a cost-effective manner for routine use in research laboratories in resource-limited settings.

American Psychological Association (APA)

Deepachandi, Bhagya& Weerasinghe, Sudath& Andrahennadi, Thisira Priyantha& Karunaweera, Nadira D.& Wickramarachchi, Nadeeja& Siriwardana, Yamuna…[et al.]. 2020. Quantification of Soluble or Insoluble Fractions of Leishmania Parasite Proteins in Microvolume Applications: A Simplification to Standard Lowry Assay. International Journal of Analytical Chemistry،Vol. 2020, no. 2020, pp.1-8.
https://search.emarefa.net/detail/BIM-1167677

Modern Language Association (MLA)

Siriwardana, Yamuna…[et al.]. Quantification of Soluble or Insoluble Fractions of Leishmania Parasite Proteins in Microvolume Applications: A Simplification to Standard Lowry Assay. International Journal of Analytical Chemistry No. 2020 (2020), pp.1-8.
https://search.emarefa.net/detail/BIM-1167677

American Medical Association (AMA)

Deepachandi, Bhagya& Weerasinghe, Sudath& Andrahennadi, Thisira Priyantha& Karunaweera, Nadira D.& Wickramarachchi, Nadeeja& Siriwardana, Yamuna…[et al.]. Quantification of Soluble or Insoluble Fractions of Leishmania Parasite Proteins in Microvolume Applications: A Simplification to Standard Lowry Assay. International Journal of Analytical Chemistry. 2020. Vol. 2020, no. 2020, pp.1-8.
https://search.emarefa.net/detail/BIM-1167677

Data Type

Journal Articles

Language

English

Notes

Includes bibliographical references

Record ID

BIM-1167677