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There are several methods for protein estimation, these are the most common:
- Near UV Absorbance (280 nm). Quantitation of the amount of protein in a solution using a simple spectrophotometer. UV detection focuses on Tyr and Trp content, and to a very small extent on the amount of Phe and disulfide bonds.
- Far UV Absorbance (205 nm). Detects various side chains, including those of Trp, Phe, Tyr, His, Cys, Met, and Arg.
These two methods uses: 0.1 M K2SO4 (pH 7.0), 5 mM KPO4 buffer (pH 7.0), Non -ionic detergent (e.g., 0.01% Brijn 35), and Guanidinium-HCl.
- The Lowry Method. This method is based on both the Biuret reaction, in wich the peptide bonds of proteins react with cooper under alkaline conditions to produce Cu+, which reacts with the Folin reagent, and the Folin-Ciocalteau reaction, where phosphomolybdotungstate is reduced to heteropolymolybdenum blue by the cooper-catalyzed oxidation of aromatic amino acids.
This methods requires: Complex-forming reagent (composed by the Solution A: 2% (w/v) Na2CO3 in dH2O; Solution B: 1% (w/v) CuSO4 5H2O in dH2O; Solution C: 2% (w/v) sodium potassium tartrate in dH2O); 2 N NaOH; Folin Reagent (commercially available); and protein standards (bovine serum albumin fraction V is widely used).
- The Bicinchoninic Acid (BCA) Assay. This method is similar to Lowry method, since it also depends on the conversion of Cu2+ to Cu+ under alkaline conditions. The Cu+ is then detected by reaction with BCA.
This methods requires: Reagent A (0.1 g sodium bicinchoninate, 2.0 g Na2CO3 H2O, 0.16 g sodium tartrate dihydrate, 0.4 g NaOH, 0.95 g NaHCO3, made up to 100 mL. Adjust pH to 11.25 with NaHCO3 or NaOH, if necessary; Reagent B (0.4 g CuSo4 5H2O in 10 mL of water). The Standard Working Reagent (SWR) is made by mixing 100 vol of A + 2 vol of B.
- The Bradford Assay. This method relies on the binding of the dye Coomassie Blue G250 to protein.
This assays requires a solution of 100 mg Coomassie Blue G250 in 50 mL of 95% ethanol, mixed with 100 mL of 85% phosphoric acid and completed to 1 L with distilled water.
These are the most widely used assays for protein estimation.
What else can I help you with?
What method are used for the estimation of molecular weight of protein?
ofcoz the protein migrate so long as it is charged ,once it become neutral it will stop migrating
What is protein estimation?
here the amount of protein in a sample can be determined, using different protocols, wherein the reagent mixture( ex: FC reagent) which when added to the sample containing protein reacts with the specific amino acids giving colour, thus the amount of protein in a sample can be estimated and the data used for further protein studies.
What are some alternatives that can be used to replace nuclear chemicals?
What do you mean by nuclear chemicals?
What is the composition of BSA?
BSA, or bovine serum albumin, is a protein derived from cows' blood. It is primarily composed of amino acids, with the main ones being alanine, arginine, aspartic acid, and glycine. BSA is commonly used in laboratory research as a blocking agent or protein standard.
What is research chemicals?
Research chemicals are ultrapure chemicals or chemicals having another special property; they are very expensive and used only in chemical research, ultratrace chemical analysis etc.
Why we are used sodium bicarbonate for the estimation of protein?
Sodium bicarbonate is not necessarily used in the estimation of protein. There are very many protein assays on the market, and the vast majority of them do NOT use NaHCO3. Look up Lowry, Biuret, Bradford, etc.
Why BSA is used as standard for protein estimation?
The Bradford assay (BSA) is commonly used for protein estimation because it is quick, sensitive, and compatible with a wide range of proteins. BSA is a reliable method for measuring protein concentration due to its colorimetric detection of the dye-protein complex, making it a popular choice in biochemical research and diagnostics.
Is lowry's protein estimation an invasive method?
No, Lowry's protein estimation method is not invasive. It is a biochemical method used to quantify the total protein concentration in a sample based on the reaction of proteins with specific reagents. It does not involve physical penetration or manipulation of the sample.
What method are used for the estimation of molecular weight of protein?
ofcoz the protein migrate so long as it is charged ,once it become neutral it will stop migrating
What is protein estimation?
here the amount of protein in a sample can be determined, using different protocols, wherein the reagent mixture( ex: FC reagent) which when added to the sample containing protein reacts with the specific amino acids giving colour, thus the amount of protein in a sample can be estimated and the data used for further protein studies.
Why antrone is used for estimation of carbohydrates?
why anthrone is used for estimation of carbohydrates?
What is the standard reference method for protein determination?
The standard reference method for protein determination is the Kjeldahl method. This method involves digesting the protein sample with concentrated sulfuric acid and then measuring the amount of nitrogen present, which is used to calculate the protein content.
Is Whey Gold Standard the best brand of whey protein?
Yes, Gold Standard is the high quality protein for you and is trusted. It is used by millions of people that want to add whey protein to their diet to make bodybuilding more effective.
What is a noun for protein?
Protein is a noun. It is any of several complex chemicals used by living things.*The similar word is the adjective protean, which has the noun form proteanism.)
How can absorbance be utilized in determining protein concentration?
Absorbance can be used to determine protein concentration by measuring the amount of light absorbed by a protein sample at a specific wavelength. This measurement is then compared to a standard curve of known protein concentrations to calculate the protein concentration of the sample.
What are the reagents used in the estimation of antioxidants?
Common reagents used in the estimation of antioxidants include DPPH (2,2-diphenyl-1-picrylhydrazyl) reagent, Folin-Ciocalteu reagent, and Trolox standard. These reagents are utilized in assays such as DPPH radical scavenging assay, total phenolic content assay, and oxygen radical absorbance capacity (ORAC) assay to measure the antioxidant capacity of compounds.
What are the standard units used to measure the concentration of a specific protein in a sample, such as in ELISA units?
The standard units used to measure the concentration of a specific protein in a sample, like in ELISA tests, are typically expressed in terms of mass per volume, such as grams per milliliter or micrograms per milliliter.
