How can the dissociation equilibrium constant, and the total number of receptors be calculated from a binding curve in a radiolabeled ligand saturation assay?

Two co-op students at your start-up company have been asked to evaluate the rate of reaction occurring in a transparent gel particle containing immobilized mouse melanoma cells. The equation for the reaction rate ris is: Vmax T'As Km+CAs where R=3.2 mm, Vmax= 0.12 gmol s' m3, CAS = 41 gmol m3, and Km=0.8 gmol m3. One student reports a reaction rate of 1.6x10 gmol st; the other reports 1.6x10 1º gmol s1. You left your calculator on the bus this morning, but must know quickly which student is correct. Use an order-of- magnitude calculation to identify the right answer. Which student is correct?

Are specific and non-specific binding determined in competitive binding assays? Or do they not have relevance?

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

An experiment was carried out to measure the reaction rate of hydrolysis of acetylcholme (substrate) with serum enzymes (Eadie, 1949). In the experiment, two experiments were conducted, namely experiment 1 without using a prostigmine inhibitor and experiment 2 using a prostigmine inhibitor at 1.5 x 10^-7 mol/l. the data obtained are:     a. Is prostigmine competitive or noncompetitive inhibitor? b. determine the value of km and rmax for the two experiments, compare

Based on your Lineweaver-Burk plot, what is the mode of PaESBL-1 inhibition by NSAMB? What is the equation for the double-reciprocal plot at 20 nMnM NAMB?

Transition-state analogs, which can be used as enzyme inhibitors and to generate catalytic antibodies, are often difficult to synthesize. Suggest a reason.

From this standard curve and chart below, does the separation of molecules in the mixture appear successful from the gel filtration? Is there a clearlydefined separation between molecules? Explain your conclusions.   Parameters required for calculation of coefficient (Kd) for unknown protein   Volume eluted (mL) Which variable does this volume represent in the equation for Kd? Fraction with maximal DNP-Aspartate detected 36 Vt Fraction with maximal Protein detected 24 Ve Fraction with maximal Blue dextran detected 6 Vo

Monoclonal antibodies can be conjugated to an insoluble support by chemical methods. Explain how these antibody-bound beads can be exploited for protein purification.

The dissociation equilibrium constant (Kd) is:_____ (select all that apply) Group of answer choices: Is a measure of ligand efficacy Typically equivalent to 1- Ka The concentration of ligand that gives maximal receptor occupancy The concentration of ligand that gives half-maximal receptor occupancy Measured in 1/M or M-1 Typically measured in mol/L or M A measure of the strength of a ligand - receptor binding reaction