Hello. I’m trying to develop adhesion assays on different cells lines. For now, I use these coatings : BSA 1% heat-denaturated, plastic, gelatin 1% (incubator 1 hour) and I fix cells with methanol. However, I notice there are "lines" or "filaments" which trap cells. This increases the absorbance and distorts my results. Have you ever noticed this problem ? According to you what is the origin of these "filaments" ? Thank you for your help.
Hi: First, you need to make sure your culture has no contamination. If your culture is OK, the filaments may be caused by methanol fixation. Methanol has the ability to denature proteins. If your coating composes of BSA and gelatin, during methanol fixation, their structures may be changed by methanol. If it is possible, you may try 4% PFA/ PBS for 10 min at RT to replace methanol.
Hi, as already sad by Chao Hsu-Wen try to take formaldehyde ( PBS buffered,I would suggest 1% @ 4°C) if it is suitable for your readout
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Biological Techniques