The quantity of my gold conjugate is too little (50uL at 20OD) to strip the membrane using the BioDot sprayers I have available (that requires volumes >1mL). Does anyone have any tips on how I can strip my membranes in order to get clean straight, and more importantly, publishable lines?
I’m afraid this is virtually impossible without the correct equipment. Any attempt to use a microsyringe and needle is unlikely to give great results. Another possibility is to use a peristaltic pump with tubing of extremely fine bore, connected to a needle. This will give you good control over flow, but you’d still need some way of moving the membrane under the needle at constant speed. With your volume, however, you are going to struggle to get lines whichever approach you use. The best option I think is to dispense 0.5 microlitre spots from a P2 pipette onto pre-cut strips. Although you don’t get a line, it doesn’t change the result of the test. You’ll just get a spot (or no spot).
One tip for lateral flow striping without a BioDot is to use a micropipette to manually apply the gold conjugate solution to the membrane. This can be done by carefully dispensing small droplets of the solution onto the membrane in the desired location. Another option is to use a small paintbrush or pipette tip to spread the solution onto the membrane. It may be necessary to practice this technique to achieve a consistent and straight line. Additionally, you may need to use a higher concentration of your gold conjugate to achieve visible results with a smaller volume. It may also be worth considering using a different detection method that does not require the use of a BioDot, such as a lateral flow assay with a lateral flow reader.
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