How do you do a Lowry assay?
- Dilute samples to an estimated 0.025-0.25 mg/ml with buffer.
- Prepare a reference of 400 microliters buffer.
- Add 400 microliters of 2x Lowry concentrate, mix thoroughly, incubate at room temp.
- Add 200 microliters 0.2 N Folin reagent very quickly, and vortex immediately.
Which method is more sensitive for protein estimation?
Compared to other methods, the BCA assay is one of the most sensitive (it can detect proteins at concentrations as low as 5 ug/mL). It has less variability than others (i.e., Bradford assay), and it can be used to measure a wide range of protein concentration.
What is the principle of BCA protein assay?
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 is Lowry’s method used to estimate protein?
Estimation of Protein by Lowry’s method Dr. Mahesha H B., Yuvaraja’s College, Mysore . Aim: To estimate the protein using Lowry’s method. Principle: The –CO-NH- bond (peptide) in polypeptide chain reacts with copper sulphate in an alkaline medium to give a blue colored complex.
What is the protocol for the Lowry assay?
PROTEINS (LOWRY) PROTOCOL 1. The “Lowry Assay: Protein by Folin Reaction” (Lowry et al., 1951) has been the most widely used method to estimate the amount of proteins (already in solution or easily-soluble in dilute alkali) in biological samples.
What is the purpose of the Lowry method?
Lowry method is one of the important methods of protein assay for determining the total amount of protein present in a given sample. So, the basic objective of this test is to estimate the amount of protein present in a sample by Lowry’s method.
Can a Lowry assay be scaled up for larger cuvettes?
As with most assays, the Lowry can be scaled up for larger cuvette sizes, however more protein is consumed. Proteins with an abnormally high or low percentage of tyrosine, tryptophan, or cysteine residues will give high or low errors]