Quantification of total and phosphorylated STAT3 by calibrated western blotting.

Quantification of intracellular proteins is essential to understand signaling. Here, we describe quantification of the expression and phosphorylation of the transcription factor STAT3. We present isolation of total and phosphorylated STAT3 from cell lysates by immunoprecipitation, followed by SDS-PAGE and western blot together with known amounts of a calibrator protein that shares an epitope with the precipitated proteins.

Quantification of membrane-bound cytokine receptors by calibrated flow cytometry.

We present a protocol for quantifying the expression of the receptor gp130 using a calibrated flow cytometric approach. We describe pitfalls for receptor quantification such as titration of primary antibodies and standardizing cell culture. Receptors are stained with primary antibodies and fluorophore-coupled secondary antibodies.

Monitor unit verification for Varian TrueBeam VMAT plans using Monte Carlo calculations and phase space data.

To use the open-source Monte Carlo (MC) software calculations for TPS monitor unit verification of VMAT plans, delivered with the Varian TrueBeam linear accelerator, and compare the results with a commercial software product, following the guidelines set in AAPM Task Group 219. The TrueBeam is modeled in EGSnrc using the Varian-provided phase-space files. Thirteen VMAT TrueBeam treatment plans representing various anatomical regions were evaluated, comprising 37 treatment arcs.

Remote EPID-based dosimetric auditing using DVH patient dose analysis.

The aim of this work was to develop and validate a method for remote dosimetric auditing that enables dose-volume histogram parameter comparisons of measured and planned dose in the patient CT volume.. The method is derived by adapting and combining a remote electronic portal imaging (EPID) based auditing method (Virtual Epid based Standard Phantom Audit-VESPA) and a method to estimate 3D in-patient dose distributions from planar dosimetric measurements.