how to do bca protein assay

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. Principle. BCA serves the purpose of the Folin reagent in the ...

When using the enhanced tube protocol (i.e., incubating at 60°C for 30 minutes), the working range for the assay shifts to 5-250 μg/mL, facilitating detection of more dilute samples. Figure 5. BCA Protein Assay Kit protocol. Rapid Gold BCA Protein Assay Kit protocol

How does the Pierce BCA Protein Assay work? The BCA Protein Assay combines the well-known reduction of Cu2+ to Cu1+ by protein in an alkaline medium with the highly sensitive and selective colorimetric detection of the cuprous cation (Cu1+) by bicinchoninic acid (BCA). What is the wavelength of BCA? 562 nm

This applies for responses producing absorbance intensities (at 562 nm) higher than 0.1. Therefore, we propose that the assay can be applied at shorter incubation times (15 min) than those indicated in manufactures specifications, and usually used by researches and industry (30 min at 37 °C).

Absorbance assay is used for determination of protein concentration. How many questions are there in BCA exam? Applicants are given sixty minutes without a calculator to complete the 40-question test. In scoring, two points are given for each correct answer and one-half point is deducted for each incorrect answer.

Based on what kit you have (I am assuming BCA) you should find information on how to do a standard curve (by making a series of dilutions of a known conc. standard protein), and what volume of BCA...

This assay proceeds in two steps: the first being the reduction of Cu 2 + by antibodies in a basic environment and the second step involves the reduced Cu + chelating with two molecules of bicinchoninic acid. 1. Prepare standard concentrations of detection antibody (Ab 2) with concentrations ranging from 1.25 to 20 μg/mL. 2.

The Pierce BCA Protein Assay is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein. This method combines the well-known reduction of Cu +2 to Cu +1 by protein in an

The BCA Assay is actually a two step reaction. 1) The first step is the biuret's reaction. Here, Cupric ions (Cu++) from cupric sulphate (or any other source of cupric) are reduced to cuprous ions (Cu+) by the amino groups on the backbone of protein chains. 2) The second step is where the BCA - bicinchoninic acid(image) comes to play.

That very simple, you need to use microsoft excel. In simple terms you need to see the equation Y=mX+C, which can be generated by plotting the std conc in Xaxis and absorbance in Y. You will get a...

7) After mixing, add 180 µL of the BCA Reagent mixture to each well of the 96 well plate. 8) Place in an incubator at 37 C for 30 min. Remove and let it cool to room temperature. Meanwhile, set the plate reader to read absorption at 560 nm. 9) After reaching room temperature, place the plate onto the plate reader and measure absorption at 560 nm

A protein assay, such the BCA Protein Assay, is an excellent tool for estimating the protein concentration of a sample. 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 Standard BCA Protein Assay enables the measurement of protein concentrations from 10 μg/mL to 2000 μg/mL with incubation at 37°C for 30 min or room temperature for 2 hours. The BCA color formation, triggered by copper reduction, is not an endpoint reaction but adds flexibility to the assay.

The Micro BCA Protein Assay Kit uses concentrated reagents and a protocol that utilizes an extended incubation time at an elevated temperature (60°C, Test Tube Procedure only). The result is an extremely sensitive colorimetric protein assay in a test tube or microplate assay format.

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. This method combines the well-known reduction of Cu+2 to Cu+1 by protein in an alkaline medium (the

The BCA protein assay is used for quantitation of total protein in a sample. The principle of this method is that proteins can reduce Cu+2 to Cu+1 in an alkaline solution (the biuret reaction) and result in a purple color formation by bicinchoninic acid. How long does it take to do a BCA assay? Choose the right BCA protein assay for your sample

BCA Protein Concentration Assay. Objective. Utilizing Beer's law, known concentrations of bovine serum albumin (BSA) are reacted with bicinchonic acid (BCA) to create a standard curve of absorbance vs. concentration of protein. This curve is then utilized to determine the concentration of protein in an unknown solution.

The BCA Protein Assay combines the well-known reduction of Cu 2+ to Cu 1+ by protein in an alkaline medium with the highly sensitive and selective colorimetric detection of the cuprous cation (Cu 1+) by bicinchoninic acid (BCA). The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex.

Biochemical analysis of proteins relies on accurate quantitation of protein concentration. This unit describes how to perform commonly used protein assays, e.g., Lowry, Bradford, BCA, and UV spectroscopic protein assays. The primary focus of the unit is assay selection, emphasizing sample and buffer compatibility.

This method involves a two-step process - (1) the reduction of cupric ions by proteins under alkaline conditions (Biuret reaction), and (2) the chelation of BCA with the Cu + ion. The second reaction produces a stable, water-soluble BCA/copper complex which absorbs light at 562 nm and changes the color of the sample from blue to intense purple.

Store at RT BCA Protein Assay Kit 1 Kit (960 assays) 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 Invitrogen Qubit Protein Broad Range Assay simplifies protein quantitation—no dilutions needed.https://www.thermofisher.com/order/catalog/product/A51292I...

The BCA protein assay is used to quantify total protein in a biological sample. BCA stands for Bicinchoninic acid, which is the key reagent used to produce a colored product.The purple colored product is analyzed in reference to a standard curve in order to quantify protein concentration. It is important to measure protein concentration after performing a protein extraction or purification ...

BCA Principle: The Pierce BCA Protein Assay is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein. This method combines the well-known reduction of Cu +2 to Cu +1 by protein in an alkaline medium ...

The BCA assay (bicinchoninic acid assay) developed by Paul K. Smith et al. has become one of the most widely used protein assays since its introduction in 1985. The advantages offered by the assay help to explain the popularity and widespread use over other protein assays. The advantages of the BCA include:

Description; Overview: The BCA protein assay is based on a biuret reaction, which is the reduction of Cu 2+ to Cu + by proteins in an alkaline solution with concentration-dependent detection of the monovalent copper ions. Bicinchoninic acid is a chromogenic reagent that chelates the reduced copper, producing a purple complex with strong absorbance at 562 nm (Smith 1985, Wiechelman 1988).

The bicinchoninic acid assay ( BCA assay ), also known as the Smith assay, after its inventor, Paul K. Smith at the Pierce Chemical Company, is a biochemical assay for determining the total concentration of protein in a solution (0.5 μg/mL to 1.5 mg/mL), similar to Lowry protein assay, Bradford protein assay or biuret reagent.

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