In the present study, we quantified the GPER binding potencies of 12 PBDEs (with bromination numbers from 1 to 8) and 18 OH-PBDEs (with hydroxyl positions of ortho-, meta- and para-) using a fluorescence
competitive binding assay in a human breast cancer cell line (SKBR3).
Competitive Binding Assay.
Competitive binding assays were performed using h[A.sub.1]AR, h[A.sub.2A]AR, or h[A.sub.3]AR expressing cell membranes (18.5 ng/[micro]L, 16.7 ng/[micro]L, or 1.7 ng/[micro]L final protein concentration, resp.) and 1.7 nM [[sup.3]H]DPCPX ([K.sub.D] = 1.7 nM), 50 nM [[sup.3]H]CGS21680 ([K.sub.D] = 23 nM), or 0.4 nM [[sup.125]I]AB-MECA ([K.sub.D] = 0.78 nM) as the respective radioligands (all purchased from PerkinElmer, Inc.
For
competitive binding assay, 2 kinds of antigen-bound plate were prepared: sLeX-acetyl phenylenediaminel (APD)-human serum albumin (HSA) and antigens derived from colo205 cells.
Binding to PPAR-a, PPAR-p/5 and PPAR-y was assessed using a
competitive binding assay, with the study finding LBE had affinity for and activated the three PPARs at 0.6, 0.4 and 1.2 pg/ mL concentrations.
Following the saturation assay, a set of
competitive binding assay were performed in membrane expressing n opioid receptors, 5 opioid receptors and k opioid receptors to determine the binding affinity of mitragynine, using the tracer [[sup.3]H]diphrenorphin and [[sup.3]H]Naltrindole which is a nonselective opioid ligand.
Development of an ELISA-based
competitive binding assay for the analysis of drug concentration and antidrug antibody levels in patients receiving adalimumab or infliximab.
PPAR[gamma]
competitive binding assay. A detailed description of the PPAR[gamma] binding assay is provided in Supplemental Material, "PPAR[gamma]
Competitive Binding Assay and Quality Assurance/Quality Control." Briefly, we used a commercially available high-throughput ligand binding assay (PolarScreen[TM] PPAR[gamma]-Competitor Assay Kit; Invitrogen, Carlsbad, CA) to investigate the binding potency of the tested compounds to PPAR[gamma] LBD.
Following the saturation assay, a set of
competitive binding assay were performed in membrane expressing [mu] opioid receptors, [delta] opioid receptors and [kappa] opioid receptors to determine the binding affinity of mitragynine, using the tracer [3H]diphrenorphin and [[sup.3]H]Naltrindole which is a nonselective opioid ligand.
Therefore, we performed a
competitive binding assay between [.sup.14]C-ATR and GHRF (Figure 4).
The vitamin [B.sub.12] content of the sequestered holo-TC is released under reducing and alkaline conditions, converted to the stable cyano form with potassium cyanide, and quantified in a
competitive binding assay with [[sup.57]Co]-cobalamin as tracer and immobilized IF as the vitamin [B.sub.12]-binding protein.
The Cbl content of the sequestered holoTC is released under reducing and alkaline conditions, converted to the stable cyano form with potassium cyanide, and quantified in a
competitive binding assay with [[sup.57]Co]Cbl as tracer and immobilized intrinsic factor as the Cbl-binding protein.
In order to facilitate comparison with the RBA, we developed the RIA to be a
competitive binding assay, run in solution with separation by filtration.
