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Reducing agent and detergent compatible protein assay rc dc assay

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The Reducing Agent and Detergent compatible protein Assay (RC/DC) is a colorimetric assay that can measure protein concentration in the presence of reducing agents and detergents. It is designed to be compatible with samples containing compounds that interfere with other protein assays.

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Lab products found in correlation

Puromycin, by Jackson ImmunoResearch (2 mentions) Ubiquitin, by Cell Signaling Technology (2 mentions) Odyssey fc, by LI COR (2 mentions) Odyssey blocking buffer tbs, by LI COR (2 mentions)

Close spelling variants of this product

DC Protein Assay (2685 citations) Bio-Rad protein assay (2654 citations) DC assay (217 citations) Detergent-compatible protein assay (76 citations) RC/DC assay (34 citations) Reducing agent (15 citations) DC (Detergent Compatible) protein assay (13 citations) Detergent compatible assay (12 citations) RC DC (11 citations) DC (detergent compatible) assay (5 citations)

2 protocols using reducing agent and detergent compatible protein assay rc dc assay

1

Quantifying Puromycin and Ubiquitin Levels

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Cells were collected as described above. Protein concentrations were measured using Reducing Agent and Detergent compatible protein Assay (RC/DC) assay (Bio-Rad, Hercules, CA, USA, 500–0119). A total of 40 μg of protein was loaded and ran on an SDS-PAGE gel, transferred to a PVDF membrane, and blocked in 5% milk in Tris-buffered saline (TBS). Membranes were probed overnight for primary antibodies specific to puromycin (Jackson ImmunoResearch, 115-035-206) and ubiquitin (Cell Signaling, 3933). Antibodies were diluted by using Odyssey Blocking Buffer TBS (LI-COR, 927-50000). LI-COR secondary antibodies conjugated with InfraRed were used according to the manufacturer’s protocol (LI-COR, 925-32211). Membranes were imaged on LI-COR Odyssey FC using IR detection. Entire lanes were normalized to the 45 kDa actin band of Ponceau S strain as a loading control. Ponceau S band densities did not differ between any groups. This was used was as a relative measure of the amount of actively translated proteins in the polysome prior to harvest of cells.
Morena da Silva F., Rosa-Caldwell M.E., Schrems E.R., Martinez L., Amos M.G., Lim S., Cabrera A.R., Brown J.L., Washington T.A, & Greene N.P. (2022). PGC-1α overexpression is not sufficient to mitigate cancer cachexia in either male or female mice. Applied physiology, nutrition, and metabolism = Physiologie appliquee, nutrition et metabolisme, 47(9), 933-948.
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2

Quantifying Puromycin and Ubiquitin Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were collected as described above. Protein concentrations were measured using Reducing Agent and Detergent compatible protein Assay (RC/DC) assay (Bio-Rad, Hercules, CA, USA, 500–0119). A total of 40 μg of protein was loaded and ran on an SDS-PAGE gel, transferred to a PVDF membrane, and blocked in 5% milk in Tris-buffered saline (TBS). Membranes were probed overnight for primary antibodies specific to puromycin (Jackson ImmunoResearch, 115-035-206) and ubiquitin (Cell Signaling, 3933). Antibodies were diluted by using Odyssey Blocking Buffer TBS (LI-COR, 927-50000). LI-COR secondary antibodies conjugated with InfraRed were used according to the manufacturer’s protocol (LI-COR, 925-32211). Membranes were imaged on LI-COR Odyssey FC using IR detection. Entire lanes were normalized to the 45 kDa actin band of Ponceau S strain as a loading control. Ponceau S band densities did not differ between any groups. This was used was as a relative measure of the amount of actively translated proteins in the polysome prior to harvest of cells.
Morena da Silva F., Rosa-Caldwell M.E., Schrems E.R., Martinez L., Amos M.G., Lim S., Cabrera A.R., Brown J.L., Washington T.A, & Greene N.P. (2022). PGC-1α overexpression is not sufficient to mitigate cancer cachexia in either male or female mice. Applied physiology, nutrition, and metabolism = Physiologie appliquee, nutrition et metabolisme, 47(9), 933-948.
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