Are specific and non-specific binding determined in competitive binding assays? Or do they not have relevance?
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Are specific and non-specific binding determined in competitive binding assays? Or do they not have relevance?
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- How do specific and nonspecific binding factor into calculations for a competitive binding assay to find the affinity of an agonist? For example, are they calculated before or after the assay, and does it affect the binding isotherm curve and how it is calculated?In a radiolabeled ligand assay to determine the binding affinity for an agonist, different concentrations of agonist are placed in test tubes with both a set number of receptors and radiolabeled ligand. Filters separate the bound and free radioligand, and a scintillation counter measures the number of bound radioligand. But how is the binding affinity of the agonist determined from all this?How does a competitive binding assay determine the affinity for an agonist? Specifically, how is the competition measured and the number of receptors/ligands counted?
- How might such a “competition assay” be used to determine the biological glycosaminoglycan that binds to protein E?In a radiolabeled ligand assay to determine the binding affinity for an agonist, how is the amount of bound radioligand measured after filtration?Why would a a pure protein screening approach be used to discover a small molecule inhibitor of a Kinesin instead of other methods? What is one advantage and one disadvantage of pure protein screens?
- How can the dissociation equilibrium constant, and the total number of receptors be calculated from a binding curve in a radiolabeled ligand saturation assay?You work for a company that manufactures a range of soy products. A customer has ask that you provide assurance that your processing operations inactivate at least 99% of the soybean trypsin inhibitor in your products. How would you develop an assay for soybean trypsin inhibitor and use it to test products??You are a technician in a biochemistry lab running receptor binding experiments. The target membrane-bound receptor has been partially purified from mouse, rat, and human cell lines. Using the same radioactive ligand in a saturation binding assay for each species' receptor, you generate the binding data in the table. The dependent variable, Y, is the fraction of binding sites occupied by the ligand. Ligand Y for mouse Y for rat Y for human concentration receptor receptor receptor (nM) 0.20 0.048 0.29 0.17 0.50 0.11 0.50 0.33 1.0 0.20 0.67 0.50 4.0 0.50 0.89 0.80 10 0.71 0.95 0.91 20 0.83 0.97 0.95 50 0.93 0.99 0.98 Determine the Ka for the human receptor in this binding experiment. human receptor Kd nM
- How would you determine the concentration of a protein that falls outside (above or below) the linear range of this assay? asapIn the "biochemical assay of b-galactosidase activity", what is o-nitrophenyl-b-D-galactosidase (ONPG) used for? O When ONPG is cleaved by b-galactosidase, we are able to assay b-galactosidase activity. O It provides essential nutrients for bacterial growth. It partially disrupts the cell membrane to allow cellular proteins to diffuse out of the cell. O ONPG cleaves the b-galactosidase that is made by the lac operon so we can see how much activity there is in the cell. ONPG cleaves galactose so we can measure how much lactose there is in the cell.Mucolipidosis type X is a lysosomal storage disease that was suggested to be a result of impaired mannose 6-phosphate receptor function. How would you test that? I need an example of a quantitative assay.