What is the proper way to dispose of blebbistatin residues?

how to prepare a BSA stock solution, then how to dilute it to 10%? and sould i dilute it in PBS or water ?

I have a problem with water solubility of Blebbistatin. I have a stock of 150mM in DMSO and need final concentration of 40µM in cell culture. As soon as I add Blebbistatin into water it precipitates. I tried stepwise reduction of DMSO but it did not work even at 75% DMSO it precipitated. I tried to dilute blebbistatin with DMSO (800µM) then add it to water and that did not work too. My cells only tolerate 0.1% DMSO concentration. So, I cannot add low concentration blebbistatin into my cell culture. Please comment if you have any experience in solublizing blebbistatin for cell culture use.

I am trying to stain a membrane protein, but the staining failed for an unknown reason. After applying the secondary antibody (Alexa 647), I did the postfixation with %3 PFA + 0.05% glutaraldehyde. Can I stain the sample again now?

I'm submitting manuscript to a journal that requires 1200 dpi in my images. I have changed the dpi from 96 but when I upload the image with 1200 dpi, the image looks super blurry when I review my manuscript. If I open the image in word/paint/other the resolution is great but in the submission it looks blurry. It is the right dpi but it is impossible to see the figures. What should I do? I don't want this to be a possible reason for the journal to not accept my manuscript.

I have tried 1% BSA in PBS and it work slightly better than my previous results but did not obtain the clarity that I need. So please suggest me any appropriate techniques to remove the background noise completely as posiible.

Hello everyone! I have started a primary cell culture from murine heart, bladder and tight. I need to do a proliferation assay using EdU-Click kit, and I was wondering how long should I incubate the cells with the EdU. I optimized the protocol on AsPc-1 (pancreatic cancer) and PS-1 ( satellite cells) cell lines, and incubation of 1h was fine, as the cells replicate quite fast. But with primary cells I guess the situation is a bit different, they replicate slower, so I guess 1h is not enough. Has someone used Edu kit with primary cells? Do you have any suggestions?

Thank you in advance!

I have intricate mouse mammary tissue organoids that have grown in 3mg/ml rat tail collagen. I would like to do immunofluorescence staining with some specific antibody. I have tried incubating the gels for upto 5 days in primary antibody and 2 days in secondary antibody. But the antibodies don't seem to penetrate through the dense collagen gel. I have also tried sectioning the gel with the vibritome and cryostat but without much success.

Any one has a bright aide about how to increase antibody accessibility?

How long (maximum) do you keep your BSA dilution at 4ºC?

I culture Endothelial cell on Polyacrylamide gel and now want to do IFA on them. However, every I fix them with 4%PFA, the cells detach and loss. Moreover, when I add coverslips after adding the mounting medium, there are so many bubble, when it's not appeared in normal cells. The last, I incubate the 1st Ab overnight at 4 and add 2nd Ab for 2h at RT but the signal is always very weak like it cant attach well. This problem is not occured on cells on object glass, only on Polyacrylamide gel. Canyone solve this or give me suggestions? Thank you in advanced, will appreciate all of answer.

regards,

vicki