Drugs that block BC cell growth or angiogenesis can restrict tumor growth and are of clinical relevance. Molecules that can target both BC cell and VEC growth as well as BC secretome may be more effective in treating After 4 or 10 h, the Ti sheets were rinsed with plenty of distilled water for 30 s to remove unabsorbed BSA, and then were incubated within 5 wt% SDS at 37C with gently shaking for 1 h to desorb the BSA. The Pierce BCA Protein Assay produces a linear response curve (R2 > 0.95) and can be performed using two different formats based upon the dynamic range needed to detect the protein concentration of an unknown sample. colorimetric detection and quantitation of total protein. Micro BCA microplate working range is from 2 - 40 ug/ml. The Micro Bicinchoninic Acid (BCA) Protei n Assay primarily relies on two reactions. (2) Accurate and sensitive, the reagent has good stability, the protein measurement range of BCA reagent is 20-200g/ml, and the micro BCA measurement range is 0.5-10g/ml. The BCA color formation, triggered by copper reduction, is not an endpoint reaction but adds flexibility to the assay. 2.7. Step 1: Prepare several dilutions of the BSA standard, at least 5. The BCA Protein Assay is a popular method for colorimetric detection and quantitation of total protein. The BCA protein assay is based on a biuretreaction, which is the reduction of Cu 2+ toCu + by proteins in an alkaline solution withconcentration-dependent detection of themonovalent copper ions. Western blots which are known as a protein immunoblot, are performed to determine the molecular weight of a targeted protein and to estimate relative amounts of the protein present in different samples.. Principle BCA serves the purpose of the Folin reagent in the Lowry assay, namely to react with complexes between copper ions and peptide bonds to produce a purple end product. Assay Purpose Measure protein concentration in solution Incubation conditions 60 min at 60C Assay Principle Micro BCA Protein Assay Kit is a ready-to-use, detergent-compatible, Pierce BCA Protein Assays have a unique advantage over the Coomassie dyebased colorimetric detection and quantitation of total protein. the Bicinchoninic Acid (BCA) Protein Assay for dilute protein samples (0.5 -20g/ml). detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein. About This Assay Caymans Micro BCA Protein Assay Kit is a microplate-based, colorimetric method for rapid total protein quantification. Bicinchoninic acidis a chromogenic reagent that chelates thereduced copper, producing a purple complexwith strong absorbance at 562 nm (Smith1985, Wiechelman 1988). The Pierce BCA Protein Assays combines the well-known reduction of Cu2+ to Cu+ by protein in an alkaline medium In conjunction with the micro-volume capability of a Thermo Scientific NanoDrop spectro-photometer, the assay provides an accurate means of Under conditions similar to the "micro BCA" assay, we determined the limit of detection for three common proteins (insulin, bovine serum albumin, and bovine gamma globulin) and found that they agree well with the existing spectroscopic detection methods. Mechanism. A stock BCA solution contains the following ingredients in a highly alkaline solution with a pH 11.25: bicinchoninic acid, sodium carbonate, sodium bicarbonate, sodium tartrate, and copper(II) sulfate pentahydrate. The BCA assay primarily relies on two reactions. First, the peptide bonds in protein reduce Cu 2+ ions The result is an Explore our BCA assay kits, that provide high-precision, detergent compatible protein quantification determination. The BCA is used for the same reasons the Lowry is used. Stoscheck (1990) has suggested that the BCA assay will replace the Lowry because it requires a single step, and the color reagent is stable under alkaline conditions. Both a standard assay for concentrated proteins and a micro assay for dilute protein solutions are described below. (B) MTT assay indicated that 500 impulses of 10 kV significantly augmented BMSC proliferation within 48 h after ESW treatment. 23208 Pre-Diluted Protein Assay Standards: Bovine Serum Albumin (BSA) Set, 7 3.5mL 23212 Bovine Gamma Globulin Standard, 2mg/mL, 10 1mL ampules 23213 Pre-Diluted Protein Assay Standards, (BGG) Set, 7 3.5mL aliquots 23246 Pierce Detergent Compatible (Bradford) Assay Kit 23235 Pierce Micro BCA Protein Assay Kit The BCA/copper complex is water-soluble and exhibits a linear absorbance at 562 nm over a broad range of protein concentrations. When it is necessary to BCA Assay: Chemical formula of Bicinchoninic acid: (HO 2CC 9H5N) 2 Molecular weight of Bicinchoninic acid: 344.33 g/mole The BCA assay is a reduction assay of Cu 2+ to Cu 1+ in For example, the dilutions may be 5, 10, 25, 50, 75, and 100 micrograms of BSA per milliliter. The Micro BCA Protein Assay Reagent Kit uses concentrated reagents and a protocol that utilizes an extended incubation time at an elevated temperature (60C). 23225), the Micro BCA Kit has been optimized for use The result is an (3) Economical and practical, in addition to test tubes, the measurement can be carried out in the microplate wells, which greatly saves the amount of samples and reagents; You add sample:reagent in 1:1 ratio which is more desirable as opposed to Pierce BCA assay. The Micro BCA Protein Assay Kit is a specialized version of the popular Pierce BCA Protein Assay for determining the protein concentration of dilute samples. You do have to incubate it for 2 hrs. The Micro BCA Protein Assay Kit is a ready to use, detergent-compatible, Western blot related, total protein analysis reagent used for the quick determination of total protein concentration of dilute protein solutions by measuring absorbance at 562 nm and comparing to a protein standard absorbance vs. concentration curve.Accurately quantifying total protein concentration is a key An adaptation of the Thermo Scientific BCA Protein Assay Kit (Product No. 23225), the Micro BCA Kit has been optimized for use chelation of two molecules of BCA with one cuprous ion. The principle of this method is that proteins can reduce Cu+2 to Cu+1 in an alkaline solution (the 3.7.1.1 Principle of the study . The reaction is similar to the well-doc-umented Lowry1 assay, but with the following improvements: The reaction reaches 90% of its maximum color development within 15 minutes thereby The first step is to separate the proteins by gel electrophoresis (SDS-page), then antigen is transferred to the blotting membrane This product is sufficient for 3,200 microplate assays when mixed with Reagents MB and MC. The bicinchoninic acid (BCA) assay is a protein quantification technique that was first demonstrated by Paul K. Smith at the Pierce Chemical Company in 1985 ( Smith et al., 1985 ). on September 17, 2022. Take3 micro-volume analysis comparison between an in situ assay and one conducted in a micro-centrifuge tube using a 20:1 volume ratio of BCA working reagent to protein standard. Views: 2. The Standard BCA Protein Assay enables the measurement of protein concentrations from 10 g/mL to 2000 g/mL with incubation at 37C for 30 min or room temperature for 2 hours. The bicinchoninic acid 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 BCA Protein Assay With its extreme sensitivity, the BCA assay has been one of the most widely used protein assays in the lab. The Micro BCA Protein Assay Kit provides an ultra-sensitive method to detect and quantify protein concentration. An adaptation of the Thermo Scientific BCA Protein Assay Kit (Product No. Based on the well-known bicinchoninic acid (BCA) Mixing together the three Micro BCA reagents results in a working solution that is sufficiently concentrated to measure protein when mixed with an equal volume of sample. The cells were subsequently lysed in BioPlex lysis buffer (Bio-Rad), corrected for protein content (micro-BCA assay), and analyzed with Luminex beadbased immunoassays (BioPlex immunoassays, Bio-Rad) of phospho-AKT (Ser 473) and phospho-ERK1/2 (Thr 202 /Tyr 204 and Thr 185 /Tyr 187) according to the manufacturer's instructions. This method involves a two-step process (1) the reduction of The September issue of Research Spotlight has arrived! Categories: Research Spotlight. BCA assay principles Chemistry of BCA-based protein assays. The Micro BCA Protein Assay Kit is a specialized version of the popular Pierce BCA Protein Assay for determining the protein concentration of dilute samples.Accurately quantifying total protein concentration is a key to most experiments and is often necessary for isolation, separation, and analysis of proteins by biochemical methods. Data are presented as the mean SD from triplicate experiments (a, P > 0.05; b, P < 0.05; c, P < 0.001 as compared to control group at the same period by One-way ANOVA with StudentNewman-Keuls post hoc test; n = 6). Were happy to present 10 recent publications from scientists whom Boster has served. Description. The Micro BCA Protein Assay Kit is a specialized version of the popular Pierce BCA Protein Assay for determining the protein concentration of dilute samples. Mixing together the three Micro BCA Reagents results in a working solution that is sufficiently concentrated to measure protein when mixed with an equal volume of sample. The concentration of desorbed BSA was measured by a micro BCA protein assay kit according to manual book. The Pierce Micro BCA Reagent A (MA) is a unique alkaline tartrate-carbonate buffer. Introduction and Principle The Bio-Rad DC Protein Assay is a colorimetric assay for protein concen-tration following detergent solubilization. The BCA protein assay is used for quantitation of total protein in a sample. Disadvantages Takes time to perform an assay.The assay is photosensitive so it required consistent illumination during the assay.Colorimetric dye-based methods The Bradford assay Bradford's method is based on a blue dye (Coomassie Brilliant Blue) that binds to free amino groups in the side chain of amino acids, especially Lysine.Advantages:.The Bradford assay is Breast cancer (BC) cell secretome in the tumor microenvironment (TME) facilitates neo-angiogenesis by promoting vascular endothelial cell (VEC) growth. The principle of this technique is quite similar to the Lowry protein assay, which is one of the alternative protein quantification methods. The kit has been optimized for determining total protein content in low Firstly, the peptide Formation, triggered by copper reduction, is not an endpoint reaction but adds flexibility the. Have to incubate it for 2 hrs 23225 ), the micro bca assay principle a! 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