One disadvantage of this detergent formulation is its relative incompatibility with certain downstream applications compared to other lysis reagents. The intensity of the colored reaction product is a direct function of protein amount that can be determined by comparing its absorbance value to a standard curve. The Pierce 660-nm Protein Assay Reagent is a ready-to-use, detergent- and reducing agent-compatible reagent to quickly measure total protein concentration. Although the Pierce 660 nm Protein Assay produces a higher level of protein-to-protein variation (37%) than other assays, such as the BCA Protein Assay, the simpler single-reagent format and broader substance compatibility make the Pierce 660 nm Assay more convenient for many routine applications. Incubate for 10 min at room temperature. Micro BCA Reagent B (MB), 240 mL Micro BCA Reagent C (MC), 12 mL Albumin Standard Ampules, 2 mg/mL, 10 x 1 mL Store at room temperature. We tested at several ages, including at 7 months, an age at which we have previously observed the onset of motor deficits (), at 5 to 6 months, before the onset The Thermo Scientific Pierce BCA Protein Assay Kit is a two-component, high-precision, detergent-compatible assay reagent set to measure (A562nm) total protein concentration compared to a protein standard.Features of the BCA Protein Assay K Lactate excretion primarily occurs because glycolysis produces NADH faster than shuttles can transport electrons into The Pierce Micro BCA Protein Assay Kit is a three-component version of our BCA reagents, optimized to measure total protein concentration of dilute protein solutions (0.5 to 20 g/mL). Thermo Scientific B-PER II Bacterial Protein Extraction Reagent (2X) is an easy-to-use cell lysis reagent is a nonionic detergent-based solutions that effectively disrupt cells and solubilize native or recombinant proteins without denaturation. 23225). Micro BCA Reagent B (MB), 240 mL Micro BCA Reagent C (MC), 12 mL Albumin Standard Ampules, 2 mg/mL, 10 x 1 mL Store at room temperature. Incubate for 10 min at room temperature. Here, Wang et al. BCA is similar to the Lowry or Bradford protein assays and was first made commercially available by Pierce, which is now owned by Thermo Fisher Scientific.In the BCA assay, a protein's peptide bonds quantitatively reduce Cu 2+ to Cu 23225, Thermo scientific, MA, USA). Protein was extracted by sonication with a probe sonicator on ice and quantified by BCA assay. IDM H&S committee meetings for 2022 will be held via Microsoft Teams on the following Tuesdays at 12h30-13h30: 8 February 2022; 31 May 2022; 2 August 2022 Western blotting. Thermo Scientific B-PER II Bacterial Protein Extraction Reagent (2X) is an easy-to-use cell lysis reagent is a nonionic detergent-based solutions that effectively disrupt cells and solubilize native or recombinant proteins without denaturation. Platelet-rich plasma, platelets, platelet membrane vesicles, and PNPs were then normalized to equivalent overall protein concentration using a Pierce BCA Protein Assay Kit (Life Technologies). Although the Pierce 660 nm Protein Assay produces a higher level of protein-to-protein variation (37%) than other assays, such as the BCA Protein Assay, the simpler single-reagent format and broader substance compatibility make the Pierce 660 nm Assay more convenient for many routine applications. Related Products Pierce IP Lysis Buffer In 1985, researchers at Pierce Chemical Company, led by Paul K. Smith, published a method detailing the use of bicinchoninic acid (BCA) as a protein quantitation reagent. The Thermo Scientific Pierce BCA Protein Assay Kit is a two-component, high-precision, detergent-compatible assay reagent set to measure (A562nm) total protein concentration compared to a protein standard.Features of the BCA Protein Assay K We tested at several ages, including at 7 months, an age at which we have previously observed the onset of motor deficits (), at 5 to 6 months, before the onset BCA Protein Assay Reagent B is a component of the Pierce BCA Protein Assay Kit, a two-component, high-precision, detergent-compatible assay that is used for total protein concentration determination compared to a protein standard. Pierce Quantitative Colorimetric Peptide Assay provides modified BCA reagents for the reduction of Cu+2 to Cu+1, a proprietary chelator optimized for the quantitation of peptide mixtures, and peptide digest reference standard for use in the generation of control linear standard curves. 23215 Compat-AbleTM Protein Assay Preparation Reagent Set, sufficient reagents to pre-treat 500 samples to remove interfering substances before total protein quantitation 90036 Protein Precipitation Plates, 2 plates, for removing protein from samples before MS analysis. Data were collected from multiple replicate wells for each experiment and were normalized to protein concentration. Protein from each sample (180 g for mouse samples, 1 mg for human samples) was reduced with 5 mM dithiothreitol for 45 minutes at 58C and then alkylated with 15 mM iodoacetamide for 45 minutes at ambient temperature in the dark. BCA is similar to the Lowry or Bradford protein assays and was first made commercially available by Pierce, which is now owned by Thermo Fisher Scientific.In the BCA assay, a protein's peptide bonds quantitatively reduce Cu 2+ to Cu We performed an enzyme-linked immunosorbent assay (ELISA) assay to detect BDNF protein in cerebellar vermis from litter-matched male and female wild-type (WT) and SCA6 84Q/84Q mice at various ages. Tissue lysates were prepared by homogenizing in ice cooled Tissue protein Extraction Reagent (Prod# 78510, Thermo Scientific, MA, USA), sonicated for 5 seconds and centrifuged for 15 min at 13,000 rpm. This BCA Protein Assay Kit is the reducing agent-compatible version of our popular Thermo Scientific Pierce BCA Protein Assay. Oxygenated cancer cells excrete much of the glucose they consume as lactate, a seemingly wasteful process that has been challenging to rationalize. Western blotting. Samples were run on SDS-page gels, electrotransferred to immobilon-P membrane (Millipore, Bedford, MA). 23225, Thermo scientific, MA, USA). BCA Protein Assay Reagent B is a component of the Pierce BCA Protein Assay Kit, a two-component, high-precision, detergent-compatible assay that is used for total protein concentration determination compared to a protein standard. Tissue lysates were prepared by homogenizing in ice cooled Tissue protein Extraction Reagent (Prod# 78510, Thermo Scientific, MA, USA), sonicated for 5 seconds and centrifuged for 15 min at 13,000 rpm. Western blotting. The Qubit Protein BR Assay Kit comes with two calibration standards, Protein BR Assay Buffer, and Protein BR Assay Reagent. Tissue lysates were prepared by homogenizing in ice cooled Tissue protein Extraction Reagent (Prod# 78510, Thermo Scientific, MA, USA), sonicated for 5 seconds and centrifuged for 15 min at 13,000 rpm. A protein assay, such the BCA Protein Assay, is an excellent tool for estimating the protein concentration of a sample. Samples were run on SDS-page gels, electrotransferred to immobilon-P membrane (Millipore, Bedford, MA). EMSAPierce BCA Protein Assay BCA rely on the formation of a Cu 2 +protein complex in a basic environment, followed by reduction of the Cu 2 + to Cu + (Smith et al., 1985).The amount of Cu 2 + that is reduced is proportional to the amount of protein present in solution. Arg levels in cells and tissues were determined using the fluorometric Arg determination kit (Biovision, #K384-100). The bicinchoninic acid assay (BCA assay), also known as the Smith assay, after its inventor, Paul K. Smith at the Pierce Chemical Company, now part of Thermo Fisher Scientific, is a biochemical assay for determining the total concentration of protein in a solution (0.5 g/mL to 1.5 mg/mL), similar to Lowry protein assay, Bradford protein assay or biuret reagent. The K + concentration in cell lysates was measured by a potassium turbidimetric assay and normalized to the protein concentration in the homogenate (mmol/g protein). The bicinchoninic acid assay (BCA) is based on a simple colorimetric measurement and is the most common protein quantification assay. The concentration of proteins was measured using a BCA Protein Assay Kit (23227, Pierce) and finally the protein concentration of each sample was adjusted to the same level. The Thermo Scientific Pierce BCA Protein Assay Kit is a two-component, high-precision, detergent-compatible assay reagent set to measure (A562nm) total protein concentration compared to a protein standard.Features of the BCA Protein Assay K A protein assay, such the BCA Protein Assay, is an excellent tool for estimating the protein concentration of a sample. Simply add 150160 L of Protein BR Assay Buffer to 10 or 20 L of your sample, followed by 30 L of Protein BR Assay Reagent. We performed an enzyme-linked immunosorbent assay (ELISA) assay to detect BDNF protein in cerebellar vermis from litter-matched male and female wild-type (WT) and SCA6 84Q/84Q mice at various ages. The BCA Protein Assay is a popular method for colorimetric detection and quantitation of total protein. For samples in which protein concentration is expected to be low (<20 g/mL), it may be necessary to use an assays alternate microplate protocol or use a specialized assay, such as the Pierce Micro BCA Protein Assay, which is specifically designed for dilute samples. The intensity of the colored reaction product is a direct function of protein amount that can be determined by comparing its absorbance value to a standard curve. The bicinchoninic acid assay (BCA assay), also known as the Smith assay, after its inventor, Paul K. Smith at the Pierce Chemical Company, now part of Thermo Fisher Scientific, is a biochemical assay for determining the total concentration of protein in a solution (0.5 g/mL to 1.5 mg/mL), similar to Lowry protein assay, Bradford protein assay or biuret reagent. The Pierce 660-nm Protein Assay Reagent is a ready-to-use, detergent- and reducing agent-compatible reagent to quickly measure total protein concentration. Mass Spectrometry Sample Preparation. In 1985, researchers at Pierce Chemical Company, led by Paul K. Smith, published a method detailing the use of bicinchoninic acid (BCA) as a protein quantitation reagent. This method combines the well-known reduction of Protein quantification was performed according to Pierce BCA Protein Assay Kit (Product No. The Thermo Scientific Pierce 660 nm Protein Assay is a ready-to-use, such as the BCA Protein Assay, the simpler single-reagent format and broader substance compatibility make the Pierce 660-nm assay more convenient for many routine applications. BCA Protein Assay Reagent B is a component of the Pierce BCA Protein Assay Kit, a two-component, high-precision, detergent-compatible assay that is used for total protein concentration determination compared to a protein standard. Protein was extracted by sonication with a probe sonicator on ice and quantified by BCA assay. g for 10 min at 4 C and adjusted to equal protein concentrations following protein quantification using the Pierce BCA Protein Assay Kit (Thermo Scientific). Micro BCA Reagent B (MB), 240 mL Micro BCA Reagent C (MC), 12 mL Albumin Standard Ampules, 2 mg/mL, 10 x 1 mL Store at room temperature. Protein from whole cell lysates was quantified using a Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). Protein concentration was determined using a Pierce BCA Protein Assay Kit (Life Technologies), and a minimum of 20 g of protein lysate was used per reaction. Primary cortical neurons were preincubated with Irisin (50 ng/mL) for 1 h and -syn PFF was administered the neurons were further incubated with Irisin for 1 or 4 d. The Thermo Scientific Pierce BCA Protein Assay Kit is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein. For samples in which protein concentration is expected to be low (<20 g/mL), it may be necessary to use an assays alternate microplate protocol or use a specialized assay, such as the Pierce Micro BCA Protein Assay, which is specifically designed for dilute samples. The Pierce 660-nm Protein Assay Reagent is a ready-to-use, detergent- and reducing agent-compatible reagent to quickly measure total protein concentration. The bicinchoninic acid assay (BCA) is based on a simple colorimetric measurement and is the most common protein quantification assay. EMSAPierce BCA Protein Assay Bicinchoninic acid (BCA) assay or Smith assay is a copper-based colorimetric assay for total protein quantification. The Thermo Scientific Pierce BCA Protein Assay Kit is a two-component, high-precision, detergent-compatible assay reagent set to measure (A562nm) total protein concentration compared to a protein standard.Features of the BCA Protein Assay K We performed an enzyme-linked immunosorbent assay (ELISA) assay to detect BDNF protein in cerebellar vermis from litter-matched male and female wild-type (WT) and SCA6 84Q/84Q mice at various ages. IDM H&S committee meetings for 2022 will be held via Microsoft Teams on the following Tuesdays at 12h30-13h30: 8 February 2022; 31 May 2022; 2 August 2022 The concentration of proteins was measured using a BCA Protein Assay Kit (23227, Pierce) and finally the protein concentration of each sample was adjusted to the same level. The values were normalized to protein concentrations measured from a BCA protein assay kit (Pierce) and the data were expressed in ng/mg protein. The Thermo Scientific Pierce 660 nm Protein Assay is a ready-to-use, such as the BCA Protein Assay, the simpler single-reagent format and broader substance compatibility make the Pierce 660-nm assay more convenient for many routine applications. Incubate for 10 min at room temperature. demonstrate that proliferating cells prefer to oxidize glucose. The K + concentration in cell lysates was measured by a potassium turbidimetric assay and normalized to the protein concentration in the homogenate (mmol/g protein). Protein was extracted by sonication with a probe sonicator on ice and quantified by BCA assay. Protein concentration was determined using a Pierce BCA Protein Assay Kit (Life Technologies), and a minimum of 20 g of protein lysate was used per reaction. Protein concentrations were determined by the Pierce BCA protein assay kit (Thermo Fisher Scientific, Cat. For each cell type, 1 x 10 6 cells were pelleted by centrifugation at 2,000 x g for 5 minutes and lysed in 1 mL M-PER Reagent for 5 minutes. g for 10 min at 4 C and adjusted to equal protein concentrations following protein quantification using the Pierce BCA Protein Assay Kit (Thermo Scientific). Here, Wang et al. The Thermo Scientific Pierce BCA Protein Assay Kit is a two-component, high-precision, detergent-compatible assay reagent set to measure (A562nm) total protein concentration compared to a protein standard.Features of the BCA Protein Assay K Pierce Quantitative Colorimetric Peptide Assay provides modified BCA reagents for the reduction of Cu+2 to Cu+1, a proprietary chelator optimized for the quantitation of peptide mixtures, and peptide digest reference standard for use in the generation of control linear standard curves. 23215 Compat-AbleTM Protein Assay Preparation Reagent Set, sufficient reagents to pre-treat 500 samples to remove interfering substances before total protein quantitation 90036 Protein Precipitation Plates, 2 plates, for removing protein from samples before MS analysis. The intensity of the colored reaction product is a direct function of protein amount that can be determined by comparing its absorbance value to a standard curve. Oxygenated cancer cells excrete much of the glucose they consume as lactate, a seemingly wasteful process that has been challenging to rationalize. Platelet-rich plasma, platelets, platelet membrane vesicles, and PNPs were then normalized to equivalent overall protein concentration using a Pierce BCA Protein Assay Kit (Life Technologies). RIPA cell lysis reagent is highly effective for protein extraction from a variety of cell types because it contains three non-ionic and ionic detergents. Protein from each sample (180 g for mouse samples, 1 mg for human samples) was reduced with 5 mM dithiothreitol for 45 minutes at 58C and then alkylated with 15 mM iodoacetamide for 45 minutes at ambient temperature in the dark. g for 10 min at 4 C and adjusted to equal protein concentrations following protein quantification using the Pierce BCA Protein Assay Kit (Thermo Scientific). The bicinchoninic acid assay (BCA) is based on a simple colorimetric measurement and is the most common protein quantification assay. The Pierce Micro BCA Protein Assay Kit is a three-component version of our BCA reagents, optimized to measure total protein concentration of dilute protein solutions (0.5 to 20 g/mL). Protein quantification was performed according to Pierce BCA Protein Assay Kit (Product No. The BCA Protein Assay is a popular method for colorimetric detection and quantitation of total protein. The protein concentration in the lysates was determined using a BCA protein assay (Pierce). This method combines the well-known reduction of This method combines the well-known reduction of The Thermo Scientific Pierce BCA Protein Assay Kit is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein. For samples in which protein concentration is expected to be low (<20 g/mL), it may be necessary to use an assays alternate microplate protocol or use a specialized assay, such as the Pierce Micro BCA Protein Assay, which is specifically designed for dilute samples. Cells were harvested at 85% confluency, washed twice and collected in ice-cold PBS, and counted. BCA is similar to the Lowry or Bradford protein assays and was first made commercially available by Pierce, which is now owned by Thermo Fisher Scientific.In the BCA assay, a protein's peptide bonds quantitatively reduce Cu 2+ to Cu Arg levels in cells and tissues were determined using the fluorometric Arg determination kit (Biovision, #K384-100). The Thermo Scientific Pierce BCA Protein Assay Kit is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein. The protein concentration in the lysates was determined using a BCA protein assay (Pierce). RIPA cell lysis reagent is highly effective for protein extraction from a variety of cell types because it contains three non-ionic and ionic detergents. IDM H&S committee meetings for 2022 will be held via Microsoft Teams on the following Tuesdays at 12h30-13h30: 8 February 2022; 31 May 2022; 2 August 2022 The cocktail contains a mixture of four inhibitors of broad specificity, including sodium fluoride, sodium orthovanadate, sodium pyrophosphate and beta-glycerophosphate. One disadvantage of this detergent formulation is its relative incompatibility with certain downstream applications compared to other lysis reagents. Halt Phosphatase Inhibitor Cocktail protects phosphoproteins from serine/threonine phosphatases as well as protein tyrosine phosphatases (PTPs) during cell lysis. Halt Phosphatase Inhibitor Cocktail protects phosphoproteins from serine/threonine phosphatases as well as protein tyrosine phosphatases (PTPs) during cell lysis. The cocktail contains a mixture of four inhibitors of broad specificity, including sodium fluoride, sodium orthovanadate, sodium pyrophosphate and beta-glycerophosphate. The cocktail contains a mixture of four inhibitors of broad specificity, including sodium fluoride, sodium orthovanadate, sodium pyrophosphate and beta-glycerophosphate. Although the Pierce 660 nm Protein Assay produces a higher level of protein-to-protein variation (37%) than other assays, such as the BCA Protein Assay, the simpler single-reagent format and broader substance compatibility make the Pierce 660 nm Assay more convenient for many routine applications. The Thermo Scientific Pierce 660 nm Protein Assay is a ready-to-use, such as the BCA Protein Assay, the simpler single-reagent format and broader substance compatibility make the Pierce 660-nm assay more convenient for many routine applications. Samples were run on SDS-page gels, electrotransferred to immobilon-P membrane (Millipore, Bedford, MA). Thermo Scientific B-PER II Bacterial Protein Extraction Reagent (2X) is an easy-to-use cell lysis reagent is a nonionic detergent-based solutions that effectively disrupt cells and solubilize native or recombinant proteins without denaturation. Related Products Pierce IP Lysis Buffer 23215 Compat-AbleTM Protein Assay Preparation Reagent Set, sufficient reagents to pre-treat 500 samples to remove interfering substances before total protein quantitation 90036 Protein Precipitation Plates, 2 plates, for removing protein from samples before MS analysis. Data were collected from multiple replicate wells for each experiment and were normalized to protein concentration. The supernatant was then transferred to a new tube, and the protein concentration of the supernatant was quantified according to the Pierce BCA Protein Assay Kit (Thermo Scientific). 23225). Lactate excretion primarily occurs because glycolysis produces NADH faster than shuttles can transport electrons into Halt Phosphatase Inhibitor Cocktail protects phosphoproteins from serine/threonine phosphatases as well as protein tyrosine phosphatases (PTPs) during cell lysis. Pierce BCA Protein Assays have a unique advantage over the Coomassie dyebased assays (Bradford)they are compatible with samples that contain up to 5% surfactants (detergents), and are affected much less by protein compositional differences, providing 23225, Thermo scientific, MA, USA). Using a BCA assay, the protein concentration of the supernatant was measured. 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