Log In Sign up
The largest database of trusted experimental protocols

Modified lowry protein assay

Manufactured by Bio-Rad
Visit Supplier

The Modified Lowry Protein Assay is a colorimetric method for the quantitative determination of protein concentration in biological samples. It is based on the reaction of protein with copper in an alkaline medium and the subsequent reduction of Folin-Ciocalteu reagent, resulting in a characteristic blue color that can be measured spectrophotometrically.

Automatically generated - may contain errors

Lab products found in correlation

Cell lysis buffer, by Cell Signaling Technology (1 mentions) Infinite 200 pro, by Tecan (1 mentions) Cell lysis kit, by Bio-Rad (1 mentions) Beuthanasia d, by Merck & Co (1 mentions)

Spelling variants of this product

Bio-Rad protein assay (2654 citations) Lowry assay (92 citations) Lowry protein assay (61 citations) Modified Lowry assay (4 citations)

4 protocols using modified lowry protein assay

1

Quantifying Secreted Proteins in Cell Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols
AMs (3–4 × 106) were plated in 6-well tissue culture dishes and incubated in the presence or absence of compounds of interest for the indicated amounts of time. Then supernatants were harvested (4 ml) and centrifuged at 500 g (10 min) and 2,500 g (10 min) to yield CM. Secreted proteins were concentrated using 3 kD Amicon size exclusion filters from EMD Millipore, after an aliquot (150 µl) was kept for LDH assay. Protein concentrations were determined by the DC protein assay (modified Lowry protein assay) from Bio-Rad Laboratories. Samples containing 30 µg protein were separated by SDS-PAGE using 12% gels and then transferred overnight to nitrocellulose membranes. After blocking with 4% BSA, membranes were probed overnight with commercially available Abs directed against SOCS (titer of 1:500), phospho- and total STAT (titer of 1:1,000), and β-actin (titer of 1:10,000). After incubation with peroxidase-conjugated goat anti–rabbit (or anti–mouse) secondary Ab (titer of 1:10,000) from Cell Signaling Technology, film was developed using ECL detection from GE Healthcare. Relative band densities were determined by densitometric analysis using NIH ImageJ software, and relative band densities for experimental conditions were expressed as described in the figure legends.
Bourdonnay E., Zasłona Z., Penke L.R., Speth J.M., Schneider D.J., Przybranowski S., Swanson J.A., Mancuso P., Freeman C.M., Curtis J.L, & Peters-Golden M. (2015). Transcellular delivery of vesicular SOCS proteins from macrophages to epithelial cells blunts inflammatory signaling. The Journal of Experimental Medicine, 212(5), 729-742.
+ Open protocol
+ Expand
2

Quantification of pSTAT6 in Beas-2B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Beas-2B cells were seeded on 6-well plates at a density of 3.0 × 105 cells/well the day before treatment. Powdered prodrugs were dissolved in DMSO to obtain 5–10 µM stock solutions. Aliquots of the stocks were added to the cell cultures to reach the desired final concentrations of the prodrugs. Cultures were incubated with prodrugs for 2 h and stimulated with IL-4 or IL-13 (2 ng/mL) for 1 h. Cells were harvested, washed with ice-cold phosphate buffer saline (PBS), and lysed with cell lysis buffer (Cell Signaling, no. 9803), supplemented with phenylmethanesulfonylfluoride (PMSF) serine protease inhibitor. After incubation on ice for 10 min, cells were collected by scraping the dishes, and the resulting suspensions were centrifuged at 13200 rpm for 15 min at 4 °C. The supernatants were transferred to clean tubes, and aliquots (5 µL) were used to measure the concentration of proteins using the modified Lowry protein assay (BioRad) on a Tecan Infinite Pro 200 plate reader. Levels of pSTAT6 and total STAT6 proteins were estimated by Western blotting. Quantification of Western blotting band density was performed using ImageJ sofware.
Mandal P.K., Morlacchi P., Knight J.M., Link T.M., Lee GR I.V., Nurieva R., Singh D., Dhanik A., Kavraki L., Corry D.B., Ladbury J.E, & McMurray J.S. (2015). Targeting the Src Homology 2 (SH2) Domain of Signal Transducer and Activator of Transcription 6 (STAT6) with Cell-Permeable, Phosphatase-Stable Phosphopeptide Mimics Potently Inhibits Tyr641 Phosphorylation and Transcriptional Activity. Journal of medicinal chemistry, 58(22), 8970-8984.
+ Open protocol
+ Expand
3

Conditioned Medium Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
AMs (1 × 106 cells/mL) were plated in serum-free RPMI 1640 in 6-well tissue culture dishes and incubated in the presence or absence of compounds of interest for the indicated amounts of time. The resulting supernatants were harvested and centrifuged at 500 × g (10 min) and 2,500 × g (12 min) to yield cell and apoptotic body-free CM (19 (link)). The resulting “neat” CM and secreted proteins were concentrated using 3 kDa Amicon size exclusion filters from Millipore (Billerica, MA). Protein concentrations were determined by modified Lowry protein assay from Bio-Rad (Hercules, CA). Samples containing 20–30 μg protein were separated by SDS-PAGE using 12.5% gels and then transferred overnight to nitrocellulose membranes. Membranes were blocked with 4% BSA and probed overnight with antibodies directed against SOCS3 (titer of 1:750), phospho-STAT3 (titer of 1:1000) and β-actin (titer of 1:10,000). A secondary antibody incubation with peroxidase-conjugated goat anti-rabbit (or anti-mouse) from Cell Signaling Technology was performed, and film was developed using ECL detection from Amersham Biosciences (Piscataway, NJ). Relative band densities were determined by densitometric analysis using NIH Image J software, and were expressed as described in figure legends.
Speth J.M., Bourdonnay E., Penke L.R., Mancuso P., Moore B.B., Weinberg J.B, & Peters-Golden M. (2016). Alveolar epithelial cell-derived prostaglandin E2 serves as a request signal for macrophage secretion of suppressor of cytokine signaling 3 during innate inflammation. Journal of immunology (Baltimore, Md. : 1950), 196(12), 5112-5120.
+ Open protocol
+ Expand
4

Acute Pulmonary Inflammation and Oxidative Stress

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mice were randomly divided into nine exposure groups corresponding to the eight temporally collected PM samples plus vehicle control (n = 5-6/group) to assess acute pulmonary inflammation and ability of particles to induce pulmonary oxidative stress in vivo. Mice were exposed via oropharyngeal aspiration to 50 µg PM and sacrificed 24 h postexposure with an overdose of Beuthanasia-D (Schering-Plough). Mice were intratracheally cannulated, and lungs were lavaged as described earlier. Caudal lobe tissue from the right lung was weighed and homogenized with a cell lysis kit (Bio-Rad, Hercules, CA), and protein concentration was obtained via a modified Lowry protein assay (Bio-Rad DC). Protein levels were utilized to normalize HO-1 results found via enzyme-linked immunosorbent assay (ELISA; Enzo Life Sciences, Farmingdale, NY) in lung homogenate to determine pulmonary HO-1 protein concentration (pg/g).
Carosino C.M., Bein K.J., Plummer L.E., Castañeda A.R., Zhao Y., Wexler A.S, & Pinkerton K.E. (2015). Allergic airway inflammation is differentially exacerbated by daytime and nighttime ultrafine and submicron fine ambient particles: heme oxygenase-1 as an indicator of PM-mediated allergic inflammation. Journal of toxicology and environmental health. Part A, 78(4).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!