Author: Tucker Burgin

SOP for bonding of PDMS to NPN

I’ve written a quick SOP document reflecting the procedure that resulting in successful permanent PDMS-NPN bonding. The procedure described in this document has not been optimized and may include unnecessary or otherwise sub-optimal steps. These steps reflect the steps that WERE

Posted in Protocols

72 Hour Diffusion Test Results

I recently got some good looking results on the FLAG diffusion test over a 24 hour period. The relative concentrations of enzyme seemed to indicate that measurably more enzyme diffused from retentate to filtrate in solutions not containing constricting Anti-FLAG

Posted in NRG

FLAG Diffusion Experiment Progress Update

For context, see my previous post. Early on in the process of designing an experiment to confirm the usefulness of a CytoVu assembly for use as an assay, it was decided that the best solution for measuring the extremely low

Posted in NRG

Solutions to the Adsorption Problem

In my last post I reported findings that the bottom well of a CytoVu assembly is capable of adsorbing protein at about 0.03 ug/mL, or 0.75ng of protein. This is a fairly small value, but for the purposes of a

Posted in NRG

Returning to CytoVu Protein Adsorption

In the wake of the apparent loss of all of the phosphatase in a sample that was placed into a CytoVu well, left for 24 hours, and then checked for phosphatase activity, it’s become clear that the adsorptive qualities of

Posted in NRG

FLAG Separation Experiments with an Enzymatic Activity Assay

As I mentioned in my previous post, there are a lot of problems associated with the technique for testing the ability of a CytoVu assembly to act as an assay that I had been trying to use before. Several possible

Posted in NRG

CytoVu FLAG Separations

Since the basic protein diffusion experiments went well, I’m moving up to something that more closely resembles an actual assay application. The basic idea is this: Biotin-conjugate FLAG antibody will be bound to Avidin acrylic beads in order to make

Posted in NRG

Public Service Announcement: Fluorescent BSA in the freezer

The following is a public service announcement: I’ve just aliquoted about 40mL of fluorescent BSA and placed it in the freezer. Each aliquot contains roughly 1mL of 6mg/mL TRITC-conjugate BSA, so there’s plenty of them. If anyone needs fluorescent BSA for anything,

Posted in NRG

Attempting to Quantify Protein Loss to CytoVu Assemblies

As I mentioned in my last post, I will be performing protein diffusion tests through CytoVu assemblies (from one well to the other) and measuring the amount of protein that is still in solution when the system reaches equilibrium in

Posted in NRG

PDMS Coating Issues

I’ve performed what might be the last test of the n-Dodecyl β-D-Maltoside (“DDM”) coating. This time I washed for three minutes each sample in HBS just like in the reference, used precisely the same concentration (and type, for that matter) of

Posted in NRG
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