assay (BioVision; Catalog # K944-100). Anti-FLAG M2 affinity gel was from Sigma (A2220). 8163). Application of linker technique to trap transiently interacting protein Then, 20 g of cortex and 10 g of glomerular lysates were boiled for 3 minutes in 6X sodium dodecylsulfate loading buffer . Biogenic antimicrobial silver nanoparticles produced by fungi trigger mitochondrial membrane protein ubiquitylation, leading to removal of damaged mitochondria (mitophagy). PRODUCT NAME: Coomassie Protein Assay Reagent PRODUCT NUMBER: 1856209 LOT NUMBER: TL275816 SPECIFICATIONS: VISUAL: Clear dark brown liquid, free of particulate matter. FK506 Binding Protein 8 Peptidylprolyl Isomerase Activity Manages a Coomassie protein assay reagent. Spin-X columns were from Costar (no. The CVs for intra-assay variability performed on three days for rPf CSP were 2.41%, 0.82% and 2% and for native Pf CSP 1.52%, 0.57%, and 1.86%, respectively. Bak activation in the absence of Bid, Bim, Puma, and p53. - PDF SECTION 2: Hazards identification Classification of the substance or mixture Classification according to Regulation (EC) No 1272/2008 [CLP] OPEN Citation: Cell Death and Disease (2016) 7, e2266; doi:10.1038/cddis.2016.167 Ofcial Journal of the Cell Death Differentiation Association When mixed with a protein solution, the acidic Coomassie-dye reagent changes color from brown to blue in proportion to the amount of protein present in the sample. . Protein kinase C gain-of-function variant in Alzheimer's disease 731-1550). Akt, a downstream target for PI3K (phosphoinositide 3kinase), is mainly activated by phosphorylation of its Ser 473 and Thr 308 residues. 8163). Proliferation and apoptosis regulation by G proteincoupled estrogen PDF Certificate of Analysis Product Name: Product Number: Lot Number Coomassie Protein Assay solution (Thermo Scientific, Waltham, MA, USA, # 1856209) was used to measure protein concentrations of each lysate sample and approximately . Protein con-centrations were measured by using a Coomassie . Chromatography columns were from Bio-Rad (no. Product code 1856209 Product name Coomassie Protein Assay Reagent Company/undertaking identification 24 hour Emergency Response: 866-536-0631 301-431-8585 . gta-20). Liquid may have a green or red tint. Protein concentrations were measured by using a Coomassie protein assay kit (Thermo Scientific; #1856209). Spin-X columns were from Costar (no. SECTION 4: First aid measures . Performing the assay in either test tube or microplate format is simple: combine a small amount of protein sample with the assay reagent, mix well, incubate b riefly and measure the absorbance at . the Coomassie Protein Assay reagent (Pierce # 1856209) (Bradford 1976). The procedure was adapted from the supplier. 1856209). Assay reagent (catalogue number 1856209, Thermo Scientic). . were stained with Coomassie Brilliant Blue to evaluate. 17-0756-05). Spleens from NOD female mice were snap-frozen and homogenized in RIPA-1640 buffer (Sigma; cat. The antibody against the S6K1 C-terminus was diluted 1:2500 The gel piece from which USP11 was identified is indicated. ASSAY: BSA conc(mg/mL) net: A595 net: 125 ug/mL 0.125 250 ug/mL 0.250 500 ug/mL 0.500 750 ug/mL 0.750 After 4 h, 10 l MTT Labelling Reagent was added to each well, then the plate was incubated overnight. Total protein (50 g) was separated by SDSPAGE (12%) Coomassie protein assay reagent: Coomassie protein assay was used to measure protein concentration. (Assay Designs (SPA-846)), Hsp70 (Assay Designs (SPA-812)), and FKBP8. Homogenously mix 0.5 ml of water and 0.5 ml of reagent and use it as blank. A kit-specific solubilisation solution was added to each well before the wells were analysed at . Kathon CG/ICP. 1 L, dilute Folin reagent for colorimetric assays; replacement reagent for DC protein assay kits and RC DC protein assay kits. Antibodies used . Product code1856209 Product name Coomassie Protein Assay Reagent www.thermofisher.com Principle Routes of Exposure/ Potential Health effects . Listenpreis: The Bradford reagent is an acidified solution of Coomassie G-250; the dye is thus primarily protonated and red. 6. The serine/threonine Akt (also known as protein kinase B) is a crucial regulator of diverse cellular processes, including cellcycle progression, cell proliferation, differentiation and apoptosis. gta-20). This assay represents a powerful tool to study Parkin E3 ligase activity and the functional interplay between ubiquitylation and . the Coomassie Protein Assay reagent (Pierce . After centrifugation for 10min at 10,000g and 4C, supernatants were transferred to new centrifuge tubes on ice. The microplate Coomassie Protein Assay Kit (product number 1856209) from Thermo Fisher Scientific Inc. (USA) was used for the Bradford assay. no. The Bradford protein assay is a time-tested colorimetric assay. . Cardiolipin assays were performed following the manufacturer's protocol. Renal cortex and glomerular lysates were collected in T-PER Tissue Reagent (Thermo 78510) containing Protease Inhibitor Cocktail (Sigma P 8340) and quantified using Coomassie Protein Assay Reagent (Thermo 1856209). 1 L, dilute Folin reagent for colorimetric assays; replacement reagent for DC protein assay kits and RC DC protein assay kits 334-20), then centrifuged (10 000 g) and the protein con-centration was measured (Coomassie Protein Assay Reagent, Thermo Scientic, Waltham, MA; cat. PMBA (2-Pyridin-4-yl-methylene-beta-boswellic acid), screened from among the 21 novel series of semisynthetic analogues of -boswellic acid, is being presented as a lead compound for integrative management of KRAS mutant colorectal cancer (CRC), upon testing and analysis for its anticancerous activity on a panel of NCI-60 cancer cell lines and in vivo models of the disease. The fluorescence was measured at an excitation wavelength of 340 nm and an emission wavelength of 480 nm. Overview. EPX260-26088-91). Plasma of patients Protein concentration was determined using the Bradford microplate protocol using Coomassie Protein Assay Reagent (cat. Glioblastoma patients have high recurrence rate and poor prognosis with only 14-18 months of survival after diagnosis ( 2 ). no. The blots were visualized as . The interacting proteins were excised as 2 mm gel pieces, digested with trypsin and identified by mass spectrometry. The protein concentration of the batch of rPf CSP used in this study was 550 g/ml and further diluted to create a stock concentration of 1ng/100 l in 1X SDS-PAGE loading dye and then two-fold serial dilutions . We have developed and employed a method to trap transiently interacting protein complexes for structural studies using glycine-rich linkers to fuse interacting partners, one of which is unstructured. The protein concentration of the batch of rPf CSP used in this study was 550 mg/ml and further diluted to create a stock Product code 1856209 Product name Coomassie Protein Assay Reagent . Appl Microbiol Biotechnol. When the dye comes in contact with protein, the first electron is donated to charged groups on the protein. Equal amounts of 70 g of protein were subjected to SDS-PAGE analysis and transferred to PVDF membrane (Millipore, IPVH00010). For each sample . SDS-PAGE was carried out using 7.5 % polyacrylamide gels containing 0.1 % SDS and stained with 0.1 % Coomassie blue R-250 after . 23200 Coomassie (Bradford) Protein Assay Kit, sufficient reagents for 630 test tube assays or 3800 microplate assays . Liquid may have a green or red tint. Protein estimation of rPfCSP was performed by using the Coomassie Protein Assay Reagent (Thomas Scientific, 1856209, Swedesboro, NJ). 1856209). Reagents. For the eclosion percentages, we set up at least 12 vials of cohorts of 2-3 heterozygous males and virgin females at 25 on standard cornmeal molasses food and under 12 hr day/night cycles. Protein concentration was determined using the Coomassie Protein Assay reagent (Pierce # 1856209 . ( a) Representative Coomassie-stained gels showing anti-GFP IPs from HEK293 extracts expressing GFP-alone or GFP-SMAD7. Cardiolipin assays were performed following the manufacturer's protocol. Coomassie protein assay reagent (ThermoFisher Scientific, cat. PRODUCT NAME: Coomassie Protein Assay Reagent PRODUCT NUMBER: 1856209 LOT NUMBER: TG267988 SPECIFICATIONS: VISUAL: Clear dark brown liquid, free of particulate matter. Defects in autophagy have been associated with aging, neurodegeneration and metabolic diseases. 2.2. PMBA (1.7-29 M . Coomassie protein assay reagent is a protein analysis reagent used for the determination of total protein concentration. We use a product from ThermoFisher (Cat# 1856209). Dialysis tubing with a diameter of 14.6 mm and a molecular cutoff (MWCO) of 6-8 kDa. When mixed with a protein solution, the acidic Coomassie-dye reagent changes color from brown to blue in proportion to the amount of protein present in the sample. The mem-brane was further incubated with primary anti-bodies of appropriate dilutions. R0278) containing EDTA/EGTA (5 m m /5 m m) and protease inhibitors (Problock, Goldbio, St Louis, MO; cat. The Pierce Coomassie Plus Assay Reagent provides increased linearity and half the protein-to-protein variability of other commercial Bradford assay formulations. no. . SAFETY DATA SHEET: GHS product identifier: Other means of identification: ASSAY: BSA conc(mg/mL) net: A595 net: 125 ug/mL 0.125 250 ug/mL 0.250 500 ug/mL 0.500 750 ug/mL 0.750 # 1856209) Non-ionic detergent or methanol (for washing the ITC system) . Dialysis tubing closures (also known as clamps) with a width of at least 35 mm. . room CAS reorder name (msds in link) Manufacturer regtype ExtraINFO messages storage; CATID; 101728: ST401: 102-71-6: t407500: triethanolamine: fisher: pubchem: Corrosive 1856209) and adjusted to 500 g/ml before using a multi-plex (26-plex) assay to detect mouse cytokines and chemokines (Affymetrix, Santa Clara, CA; cat. no. 1856209). Coomassie (Bradford) Protein Assay Kit Component # Description : 0023209 . No treatment for NAFLDs is currently available, but autophagy . 5000207; BioRad Laboratories, Inc.). 8. Glioblastoma is a high-grade malignant tumor in the human brain ( 1 ). Genetic, structural, and biochemical analyses of IS607-family transposons shows that the DNA translocation reaction proceeds very differently from other reactions promoted by serine recombinases. was used in the indicated assays to allow for protein expression to be detected by . Coomassie protein assay reagent was from Thermo Scientific (no. 731-1550). with the Coomassie Protein Assay Reagent (#1856209, Thermo Scientific). We and other groups have previously reported in vitro assays of Parkin E3 ligase activity using . Protein assay and SDS-PAGE. DC protein assay Eclosion assay. of rPf CSP are described earlier [17,21]. Simply add the reagent to equal volumes of samples and standards, mix, then measure the absorbance at 595nm. The epicardium is the main source of RA in the embryonic heart, yet the cardiogenic functions of epicardial-produced RA are not fully understood. 17-0756-05). After centrifugation (20 min at 20 x g000 ), the protein concentration in supernatants was determined using Coomassie protein assay reagent (1856209, Thermo Scientific) and bovine serum albumin as standard. One of the biggest benefits of this method is that the BCA protein assay is compatible with most protein samples and protein samples that contain up to 5% surfactants (detergents). by centrifugation and soluble proteins concentration determined using Bradford assay (Thermo Scientific, 1856209). Non-alcoholic fatty liver diseases (NAFLDs) are characterized by hepatic fat accumulation with or without inflammation. Chromatography columns were from BioRad (no. SDS-PAGE was carried out using 7.5 % polyacrylamide gels containing 0.1 % SDS and stained with 0.1 % Coomassie blue R-250 after electrophoresis. The basis for the assay is that in order for the Coomassie dye to bind stably to protein, it needs to be doubly protonated. 5. The parents were transferred to a new vial every day. The Pierce Coomassie Plus Assay Reagent is a single, ready-to-use solution for measuring protein concentration. GB-334-20), then centrifuged (10 000 g) and the protein concentration was measured (Coomassie Protein Assay Reagent, Thermo Scientific . Current treatments include surgical tumor removal and radiotherapy followed by Temozolomide chemotherapy. GFP-Trap-A beads were from Chromatek (no. Protein concentration was determined using (i) BSA standards on an SDS/PAGE gel stained with Coomassie Brilliant Blue stain and (ii) A 595 of BGG standards or PKC protein after mixing 5 L of protein solution with 45 L of buffer (0.05 M NaOH and 20 mM Tris) and 500 L of Coomassie (Bradford) Protein Assay Reagent (1856209; Thermo Fisher . We utilize Coomassie Protein Assay Reagent (Thermo Scientific, 1856209), though other reagents can also be used. GFP-Trap-A beads were from Chromatek (no. 2.4. ASSAY: BSA conc(mg/mL) net: A595 net: 125 ug/mL 0.125 250 ug/mL 0.250 500 ug/mL 0.500 750 ug/mL 0.750 The fluorescence was measured at an excitation wavelength of 340 nm and an emission wavelength of 480 nm. Compare all available Bradford assays Features of the Coomassie Plus Protein Assay include: Colorimetric measure with a standard spectrophotometer or plate reader at 595 nm Samples were resolved by 10 % SDS-PAGE and transferred to a polyvinylidene difluoride (PVDF) mem-brane (Immobilon-P, Millipore). 1856209). The Pierce Coomassie Protein Assay Kit is a ready-to-use formulation of the popular assay reagent originally described by Bradford in 1976. . Glioblastoma patients have high recurrence rate and poor prognosis with only 14-18 months of survival after diagnosis ( 2 ). The Coomassie dye in the Coomassie (Bradford) reagent binds to protein in the sample leading to an immediate shift in absorption with a concomitant color change of the solution from red to blue. Here, we investigated the roles of RA signaling in the embryonic epicardium using in vivo and in vitro models of excess or deficiency of RA. CHEMTREC: 800.424.9300 Outside US: 703.527.3887. 15 g of total protein were separated by 8% sds-page, transferred to nitrocellulose, and incubated with primary antibody at the indicated dilutions overnight at 4 c in tbs + 0.1% tween 20 + 5% Kit Contents: Coomassie . 1856209; Thermo Fisher Scientific, Inc.) and the Quick Start Bovine Serum Albumin Standard Set (cat. the protein concentration in the supernatants was assessed by the bradford assay using the coomassie protein assay reagent (pierce catalog no. Protein concentration was determined using Coomassie Protein. Protein concentration was determined using (i) BSA standards on an SDS/PAGE gel stained with Coomassie Brilliant Blue stain and (ii) A 595 of BGG standards or PKC protein after mixing 5 L of protein solution with 45 L of buffer (0.05 M NaOH and 20 mM Tris) and 500 L of Coomassie (Bradford) Protein Assay Reagent (1856209; Thermo Fisher . the Bradford assay using the Coomassie Protein Assay Reagent (Pierce catalog no. 1856209). Anti-FLAG M2 affinity gel was from Sigma (A2220). The absorbance at 595 nm was measured in a Microplate Reader (Bio Tek, USA). Biotechnology, Inc.). The protein standards with molecular weights ranging from 10 to 260 kDa (Fermentas #SM1841) were used. Autophagy is a fundamental housekeeping process by which cells degrade their components to maintain homeostasis. Read "10.1016/j.mimet.2014.10.006" on DeepDyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips. reproducible; the Coefficient of Variation (CV) for inter-assay variability for rPf CSP and native PfCSP were 1.74% and 1.32%, respectively. Total protein quantification was conducted by Bradford Coomassie protein assay (#1856209; Thermo Scientific) according to standard protocols. Albumin Standard: 1856209 : Coomassie Protein Assay Reagent: Albumin Standard Not available. The assay costs only pennies per sample and can be performed in either test tube or microplate format. Cuvettes with two clear windows and a pathlength of 10 mm. Glutathione sepharose beads were from GE healthcare (no. ABF3 phosphorylation was quantified by phosphorimaging and normalized to protein amount using Coomassie brilliant blue (CBB) gel staining (lower panel). Cell viability was assessed after 72 h using an MTT assay. Product code1856209 Product name Coomassie Protein Assay Reagent www.thermofisher.com. The Role of GSK-3 in the Regulation of Protein Turnover, Myosin; Identification and Characterization of Protein Kinase C Substrates in Human Breast Cells; A Specific Inhibitor of Calcium/Calmodulin-Dependent; The Contributions of Protein Kinase a and Protein Kinase C to the Actions of 5-HT on the L-Type Ca *+ Current of the Sensory Neurons in .