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Proteins reduce alkaline Cu (II) to Cu (I) in a concentration-dependent manner. The samples were vortexed and sonicated in an ice water bath. E001 ELISA Microplate(Dismountable) 86/812 2 -8C/ 20C . BCA protein assay kit reducing agent compatible (microplate) Sample type: Cell Lysate, Purified protein, Tissue Lysate. Shipping. Add 200 l of BCA working reagent to each well and mix well. The kit has been formatted for microplate-based protein assays. TRIzol Reagent, ampicillin, chloramphenicol, glycerol, CHAPS, imidazole, Pierce TM Universal Nuclease for Cell Lysis, HisPur TM Ni-NTA Resin, Pierce TM Microplate BCA Protein Assay Kit - Reducing Agent Compatible, PureLink TM Quick Plasmid Miniprep Kit, S.O.C. This is inline with what the user manual specifies. To carry out the protein-lipid overlay assay, dried acidic lipids were reconstituted with 10 l of CHCl 3:MeOH:H 2 O (1:2:0.8). US EN. Discard any kit reagent that is discolored or appears to have microbial contamination. Add 200l Working Solution to each tube, seal and vortex to mix. Manufacturer: Thermo Scientific 23235 Catalog No. Next, 100 g of protein was mixed with 2X substrate solution (Z-DEVD-R110) and incubated for 30 min at 37C. generated by reduction of cupric cation (Cu2+) by the protein in alkaline conditions. . The current imaging AVAM Sciences is focused on manufacturing clinical and analytical instruments and diagnostic reagents. Epigenetic Kits (4) Functional Assay . The kit also includes Bovine Serum Albumin (BSA) as a reference protein standard. (BCA) Protein Assay . The proteins were separated by SDS-PAGE (4~12%) and . Protein Carbonyl Content Assay Kit ab126287 is designed to provide a simple and accurate method of quantifying carbonyls in protein samples. . Applications Products Services Support. Find microplate bca protein assay kit and related products for scientific research at MilliporeSigma. Description: The BCA Protein Assay Kit can be used to measure the protein concentration of lysates or homog-enates, in microplate format, prepared with the following buffers: Cell Lysis Buffer (10X) #9803, RIPA Buffer (10X) #9806, PathScan Sandwich ELISA Lysis Buffer (1X) #7018. The kit is shipped at ambient temperature. color formation with BCA2. 200-1000 g/ml protein. Cover plate and incubate at 37 for 2 hours or place at 60 for 1 hour. The Micro Bicinchoninic Acid (BCA) Protein Assay is a highly sensitive colorimetric assay that is compatible with detergent-solubilized protein solutions and is a modification of the Bicinchoninic. The amount of reduced copper is directly proportional to the amount of total proteins. Ships Today (1) Product Category. Originally, BCA assay was developed in test tube format and, according to the Thermo Scientific Pierce BCA Protein Kit user guide , was based on 1:8 ratio between the sample and working reagents mix (WR) volumes. 3800 microplate assays . Cover the plate and incubate at 37C for 30 minutes. The absorbance is directly proportional to protein . 5. Note: The protein concentrations of lysates were measured by a Micro BCA protein assay reagent kit following the instructions of the manufacturer. Principle of the Assay This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Amplite Colorimetric BCA Protein Quantitation Assay Kit can be performed in a convenient 96-well microtiter-plate format and easily adapted to automation with no separation steps required. The remaining kit components should be stored at room temperature. 4. Demonstration: Elaine ChengEditing: Michael CaiVoice-over: Huiying Zhang (Amelie) The BCA Protein Assay is used to determine the protein concentration of an unknown sample. The kit has been formatted for microplate-based protein assays. BCA Protein Assay kit provides a colorimetric detection and quantification of total protein content even in the presence of detergents, measuring total protein concentration of pure proteins, extracts or lysates. The assay is linear over a wide range of protein concentrations (25-2000 g/mL). The microplate is sealed and allowed to incubate at 37 C for 30 minutes. Absorbance was measured at 490 nm using a microplate reader (Infinite M1000 Pro, Tecan, Mnnedorf, Switzerland). The kit has been optimized for determining total protein content in low protein concentration samples (0.5-40 g/mL), even in the presence of detergents. The assay is linear over a wide range of protein concentrations (25-2000 g/mL). In this assay, copper (II) [Cu2+] is reduced to copper (I) [Cu1+] in the presence of peptides in alkaline solutions. BCA Protein Assay Kit II (ab287853) Microplate Method 1. Thermo Scientific Pierce Microplate BCA Protein Assay Kit - Reducing Agent Compatible Reducing agent-compatible version of our popular BCA Reagent Kit to measure protein concentration in samples containing DTT or BME thiol-reductants. +2 Cu is reduced to Cu +1 in the presence of protein when in an alkaline medium and is chelated to BCA, leading to absorbency at a wavelength of 562 nm and demonstrating linear correlation to protein values. Search Within. Bicinchoninic Acid Kit for Protein Determination, BCA. : EH4755 Revision: V 3.0 Size: 48T/96T Reactivity: Human . Subsequently, 30 g lysate proteins were subjected to each lane and . Colorectal cancer (CRC) accounts for approximately 10% of all new cancer cases worldwide with significant morbidity and mortality. Whole cell lysates were centrifuged at 13,000 rpm for 20 min at 4 C and quantified using a SMART BCA Protein Assay Kit (iNtRON). BCA protein assay kit for low concentrations (ab207002) Specific References (11) (Bradford) Protein Assay Kit is a quick and ready-to-use modification of the well-known Bradford coomassie-binding, colorimetric method for total protein quantitation. . Cool down the plate to RT. The Micro BCA Protein Assay Kit (Product No. medium, and Luria-Bertani (LB) agar were acquired from Thermo Fisher Scientific . All Luminex High Performance Assay Panels are validated for use with serum and plasma . Our products are used in many settings from laboratories to physician offices. Screening for different acute phase proteins, cytokines, and chemokines in cell lysates were performed with a Proteome profiler array (Mouse Cytokine Array Panel A Array kit) from R&D Systems [30] , [31] , [32] . Make also a duplicate or triplicate of your sample, to ensure the repeatability of the assay. Briefly, 25 L of protein standard or sample are added to the micro-well followed by the addition of 200 L BCA working reagent (1:8). and the supernatants were collected. In brief, 1 x 10 6 treated cells were lysed in lysis buffer provided in kit and centrifuged at 5000 rpm for 5 min. Add 150 myl of working solution to each well and mix thoroughly on the plate shaker for 30 seconds. Support on how to determine protein concentration can be found here. Thermo Scientific Micro BCA Protein Assay Kit Special 3-component version of our popular BCA Reagents to measure total protein concentration of dilute protein solutions (0.5 to 20 micrograms/mL). 8. Pipette working solution into microplate wells (100l/well) Add gradient volume of standard protein (2mg/ml) to each well (5 or more) followed by adding of sample protein (5l/well) Mix wells thoroughly with pipet. Learn more about the BCA assay: http://www.piercenet.com/product/bca-protein-assay.Accurately measure the total protein concentration in your sample with thi. Protein)ELISA Kit Catalogue No. Supernatant was collected and concentration of protein was determined by BCA protein estimation kit (Sigma-Aldrich, USA). standard microplate protocol: 25-1,000 g/ml assay results in 30 min at room temperature excellent linear range of 15-1,000 g/ml and sensitivity down to 15 g/ml two stable reagents used to make one working reagent less than 10 min sample prep time compatible with commonly used mass spec reagents simple, easy to perform accurate Platform Absorbance microplate reader Recommended plate Clear bottom Components Example protocol AT A GLANCE Protocol summary The Microplate Procedure affords the sample handling ease of a microplate and requires a smaller volume (10-25 L) of protein sample; however, because the sample to working reagent ratio is 1 x 96-well microplate and Magnetic antibody-coated beads: Species: Human: Analytes Detected: Please see analyte list in assay customization tool below. There are two different procedures: one using test tubes and another using microplates. . The experiments were done in at least triplicate. Protein samples were loaded onto SDS-PAGE gels and transferred . 4. The concentrations of total proteins were measured by BCA protein assay kit. AVAM Sciences is focused on manufacturing clinical and analytical instruments and diagnostic reagents. Kit Contents: BCA Reagent A, 1000mL (in Product No. A newer version of the assay, the Pierce Rapid Gold BCA Protein Assay Kit, simply requires incubation for five minutes at room temperature and yields an intense orange-gold color that is measured at 480 nm. 2.3. 8.3. This is the most popular assay type for measuring protein carbonyl content. BCA Protein Assay Kit 71285-3 Description The BCA Protein Assay is based on a biuret reaction, the reduction of Cu 2+ to Cu 1+ by protein in an alkaline solution, and . . Products Building Blocks Explorer Technical Documents Site Content Papers Genes Chromatograms. PI23252 $337.00 / Each of 1 Qty Check Availability Add to cart The unique, patented method ulizes bicinchoninic acid (BCA) as the detecon reagent for Cu, which is formed when Cu is reduced by protein in an alkaline environment.A purplecolored reacon product is . Expiry Date: see kit label Principle: Sandwich NOTE: FOR RESEARCH USE ONLY. (BCA) Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). 23225 Pierce BCA Protein Assay Kit, sufficient reagents for 500 test-tube or 5000 microplate assays . Notes This product is manufactured by BioVision, an Abcam company and was previously called K818 BCA Protein Assay Kit - Reducing Agent Compatible. Procedure 6-Microplate Procedure Prepare the Working Reagent. Our lab quantitates protein using microplates so we followed this procedure: Standards are made using diluent of sample and BSA that comes with the kit. K818-1000 is the same size as the 1000 test size of ab207003. The result is an extremely sensitive colorimetric protein assay in a test tube or microplate assay format. This reducing agent-compatible version of Thermo Scientific's Pierce BCA Protein Assay enables one to measure protein concentration in samples that contain thiol-reductants dithiothreitol (DTT) and 2-mercaptoethanol (BME). 23225) that is optimized for quantification of dilute protein samples. Our Micro BCA Protein Assay uses concentrated reagents and a protocol with extended incubation time at an elevated temperature, which results in an extremely sensitive colorimetric protein assay, enough for 480 test tube and about 3,200 microplate assay formats. Absorbance was then measured at 450 nm, with a reference wavelength of > 650 nm, on the Synergie multiwell scanning spectrophotometer (BIOKOM, Janki, Poland). Bicinchoninic acid is a highly specific chromogenic reagent for Cu (I), forming a purple complex with an absorbance maximum at 562 nm. Protein concentrations were determined using a BCA protein assay kit (PIERCE). Working reagent, clear liquid in the color of yellow green, is mixed with the Reagent A and Reagent B at ratio of 50:1. The assay is based on the reaction of bicinchoninic acid (BCA) with the cuprous cation (Cu 1+ ), which is generated by reduction of cupric cation (Cu 2+) by the protein in alkaline conditions. 23227), the Micro BCA Kit has been optimized for use with dilute protein samples (0.5-20 g/ml). Microplate reader (2) Microplate (12 x 8 well strips) (1) Product subtype Clear. 23227 Pierce BCA Protein Assay Kit, sufficient reagents for 250 test-tube or 2500 microplate assays . The BCA Protein Assay is one of the most commonly used methods for protein quantification. The kit also includes Bovine Serum Albumin (BSA) as a reference protein standard. BCA can chelate (bind to) Cu1+, creating a purple color with a maximum absorbance at 562 nm. BC Assay : Protein Assay Kit Product Description High quality reagents for the determination of protein concentration by the bicinchoninic acid method. The BCA Protein Assay Kit can be used to measure the protein concentration of lysates or homogenates, in microplate format, prepared with the following buffers: Cell Lysis Buffer (10X) #9803, RIPA Buffer (10X) #9806, PathScan Sandwich ELISA Lysis Buffer (1X) #7018. Our products are used in many settings from laboratories to physician offices. Kit Components No. The dynamic range for this assay is 0.125 - 2 mg/mL. The absorbance was measured at 570 nm by a microplate reader (BioTek, USA), and the relative cell viability (%) was expressed as a percentage relative to the control group cells (100%) at last. Features of the BCA Protein Assay Kit include: Colorimetric estimate visually or measure with a standard spectrophotometer or plate reader at 562 nm Uniformity exhibits less protein-to-protein variation than dye-binding methods (Bradford) Compatibility unaffected by typical concentrations of most ionic and non-ionic detergents It is . Use the BCA or Lowry Protein Assay A precipitate forms in all tubes : Sample contains a . Features of the BCA Protein Assay Kit include: Colorimetric estimate visually or measure with a standard spectrophotometer or plate reader at 562 nm Uniformity exhibits less protein-to-protein variation than dye-binding methods (Bradford) Compatibility unaffected by typical concentrations of most ionic and non-ionic detergents Cool plate to room temperature (RT). BCA assay. 23235) from Thermo Scientific Pierce Protein Research Products is an adaptation of the BCA Protein Assay Kit (Product No. 23225) or 500mL (in Product No. The standard assay requires 100 ul of standards or samples per tube whereas the microplate assay is far more economical, requiring only 5 ul of standard or samples per well. Add 25 l of each standard and protein samples into separate microplate wells. The bicinchoninic acid (BCA) assay is one of many ways to measure the total protein concentration of an unknown sample. The dynamic range for this assay is 0.125 - 2 mg/mL. Performance Validation: Our Luminex High Performance Assays undergo our most extensive validation testing. Upon arrival, store the protein standards at 4C. The Micro BCA Protein Assay Kit uses concentrated reagents and a protocol that utilizes an extended incubation time at an elevated temperature (60C, Test Tube Procedure only). generated by reduction of cupric cation (Cu2+) by the protein in alkaline conditions. The Pierce microplate BCA protein assay kits and the Invitrogen Qubit Protein BR Assay Kit were used with a microplate and PCR tube, respectively. The kit has been formatted for microplate-based protein assays. 27 Thermo Scientific Pierce Bovine Gamma Globulin Standard Ampules, 2 mg/mL An adaptaon of the BCA Protein Assay Kit , the Micro BCA Kit has been opmized for use with dilute protein samples (0.520g/ml). Kit Contents: Coomassie (Bradford) Protein Assay Reagent, . The Thermo Scientific Pierce BCA Protein Assay Kit is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric . Protein concentration in the reconstituted AT preparations Anbinex and Kybernin was determined with a commercially available bicinchoninic acid (BCA) assay kit (Pierce BCA Protein Assay Kit; Thermo Fisher Scientific) using a VERSAMax microplate reader with SoftMax Pro 7.0.3 software. 3. The protein concentration was determined using the Pierce protein assay reagent according to the manufacture's instruction. Capture antibody was pre-coated onto 96- Manufacturer: Thermo Scientific 23252 Catalog No. 2. Under alkaline condition, the reduction of Cu2+ to Cu+ is realized by peptide bonds in proteins (biuret reaction). The protein carbonl assay protocol is based on the reaction of DNPH with protein carbonyls. Pierce BCA Protein Assay Kit INSTRUCTIONS Pierce Biotechnology PO Box 117 (815) 968 -0747 www.thermoscientific.com/pierce 3747 N. Meridian Road Rockford, lL 61105 USA (815) 968 -7316 fax Number Description 23225 Pierce BCA Protein Assay Kit , sufficient reagents for 500 test-tube or 5000 microplate assays Advanced Search. The working reagent, based on bicinchoninic acid (BCA), is used for the colorimetric detection of protein . PI23235 $308.00 / Each of 1 Qty Check Availability Add to cart Description Specifications Documents 2. The microplate was incubated for 18 h at 37 C and 5 % CO 2. It has all of the benefits of the BCA protein assay. . Item Specifications(48T/96T) Storage E001 ELISA Microplate(Dismountable) 86/812 2 -8C/ 20C E002 Lyophilized Standard 1vial/2vial 2 -8C/ 20C E039 Sample Dilution Buffer 10ml/20ml 2-8C 23252 1997.3 Pierce Microplate BCA Protein Assay Kit - Reducing Agent Compatible Notes and symbols The Pierce Micro BCA Protein Assay Reagent Kit is a detergent-compatible bicinchoninic acid formulation for the colorimetric detection and quantitation of total protein. 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