[Spectrophotometric determination of protein content in THP-1 monocytes/macrophages - description of the method].
Proteins are the basic building block of tissue, and are part of enzymes and hormones regulating many important life processes. Changes in their concentration control the metabolic processes of the cell. Quantitative determination of the protein content is divided into indirect methods (e.g. Kjeldahl method) and direct methods (buret method, Lowry, immunoenzymatic, formol method, based on incorporation of dye in the range of ultraviolet spectrophotometry, and based on the phenomenon of selective absorption of radiation in the infrared range). One of the methods for the determination of protein content is the spectrophotometric method described by Bradford. The protein concentration assay procedure utilizes the phenomenon of formation of the dye (Coomassie Brillant Blue G-250)-protein and colour intensity is proportional to the protein content in the solution. The aim of this study was to verify the usefulness of this method for determining the protein content in THP-1 cells cultured with extracts of nettle fruit stalks (Urtica dioica L.). Aqueous and alcohol extracts at two concentrations were used. It has been shown that the spectrophotometric determination of protein content by the Bradford method is an effective and accurate method for determining the concentration of protein in THP-1 macrophages. The results indicate that this method can be recommended for the determination of the protein content in other cell cultures.
Jolanta
J
Wolska
Katarzyna
K
Janda
Izabela
I
Gutowska
D002460:Cell Line; D003124:Colorimetry / Q000379:methods; D010940:Plant Proteins / Q000032:analysis*; D018547:Plant Stems / Q000737:chemistry*; D012394:Rosaniline Dyes / Q000008:administration & dosage; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D026982:Urtica dioica
D010940:Plant Proteins; D012394:Rosaniline Dyes; C004692:coomassie Brilliant Blue