I am staining for protein x in leishmania. I have tried so many different method of fixation e.g. 1-4%PFA (10 min), methanol, formaldehyde (1-4% for 5-10min), Acetone:methanol (1:1) in all combinations of time Vs concentrations. I was not getting staining for that protein. I tried acetone for fixation (5 min/RT). Now, I am getting staining but cell size got shrunken. Can anyone suggest me protocol to improve staining. I am primarily using antibody which is working well with mammalian cells
Please give us some more information. Do you work with frozen sections? cells? paraffin sections? which species? what says the antibody-datasheet? ab raised against which species? which application produced for?
For Leishmania : Promatigotes retain their shape by fixing in situ in culture by 5 % formaldehyde. They retain shape. Try this method.
Hi G-Halli Rajasekariah,
The problem is not fixing the promastigote. Main problem is staining. I have tried all types of fixation, but I am getting staining in only Acetone fixation.
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Antibodies