Bronchoalveolar lavage fluid (BALF) was centrifuged at 4000 rpm for 10 minutes at 10°C. Supernatant was collected, aliquoted, and stored at −80°C. For analysis, samples were thawed, and protein content was determined using a Pierce 660 nm protein assay reagent kit (CAT# 22662) (Thermo-Fisher Scientific) according to the manufacturer’s protocol using a 96 well plate and a Spectramax M5 microplate reader (Molecular Devices, Sunnyvale, CA, USA) utilizing SoftMax Pro software (v6.5, Molecular Devices) set to an absorbance point measurement at 660 nm. Heme content was determined by calculating the absorbance of the BALF at 540 nm using a Spectramax M5 microplate reader, using a 96 well plate and an absorbance point measurement at 540 nm.
Pierce 660 nm protein assay reagent kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Pierce 660 nm Protein Assay Reagent Kit is a colorimetric assay used to quantify the total protein concentration in a sample. The assay reagent binds to proteins, resulting in a color change that can be measured spectrophotometrically at 660 nm. The kit provides a simple, rapid, and accurate method for determining protein concentrations in a variety of samples, including cell lysates, purified proteins, and biological fluids.
Automatically generated - may contain errors
Lab products found in correlation
Spectramax m5, by Molecular Devices (1 mentions)
Softmax pro, by Molecular Devices (1 mentions)
Nicotinamide adenine dinucleotide, by Merck Group (1 mentions)
Griess reagent, by Thermo Fisher Scientific (1 mentions)
2 7 dichlorofluorescin diacetate, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Thiobarbituric acid, by Merck Group (1 mentions)
Cytochrome c, by Merck Group (1 mentions)
Phosphoric acid, by Merck Group (1 mentions)
Pyrogallol, by Merck Group (1 mentions)
O phthalaldehyde, by Merck Group (1 mentions)
L glutathione reduced, by Merck Group (1 mentions)
2 protocols using pierce 660 nm protein assay reagent kit
1
Protein and Heme Quantification in BALF
2
Comprehensive Biochemical Assays Protocol
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Thermo Scientific Pierce 660 nm Protein Assay reagent kit was purchased from Thermo Scientific (Pierce) for protein quantification. Griess reagent was purchased from Thermofisher Scientific. 2,4‐Dinitrophenylhydrazine (DNPH) and nicotinamide adenine dinucleotide phosphate (NADPH) were purchased from Tokyo Chemical Industry America. Phosphate‐buffered saline (PBS), nicotinamide adenine dinucleotide (NADH), 2`, 7‐dichlorofluorescindiacetate (DCF‐DA), pyrogallol, phosphoric acid, O‐phthalaldehyde (OPT), L‐glutathione reduced, cytochrome c, bovine serum albumin (BSA) trichloroacetic acid and thiobarbituric acid were purchased from Sigma Aldrich.
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