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 taken to achieve a successful bond, not necessarily the steps that MUST be taken. —…
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 antibody, and this is excellent news, but over the 24 hours between beginning the diffusion…
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 concentrations [ng/mL scale] of bacterial alkaline phosphatase (“bAP”) in the filtrate which would indicate what…
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 sensitive separation such as the FLAG tests that I’ve been working on, it can be…
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 the CytoVu assemblies needs further characterization. I showed in an earlier post that there was some loss…
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 sources of error were apparent, such as the difficulty inherent in consistently pipetting such small…
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 them far too large to pass through the MP8NP50 membrane. Then, Bacterial Alkaline Phosphatase (bAP)…
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, feel free to take as much as you need from the box labeled “TRITC-conjugate BSA”…
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 order to determine the amount of protein that is adsorbed to the assembly and thus…
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 protein that was used in the reference, and tested a positive control, a negative control,…