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immunofluorescence staining of sperm cells


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#1 tola

tola

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Posted 10 March 2010 - 04:53 AM

Hi,

Having trouble in staining sperm cells that I'm taking out of the epididydmis with anti ubiquitin antibody.

for some reason the cells fell off the slides. I did methanol fixation for 15 min. and the antibody didn't stain at all although the data sheet say it is good for staining.

does any one have any tips??

something???

please

I going carzy.

thanks

tola

#2 bob1

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Posted 10 March 2010 - 03:28 PM

Try using APS coated slides or coating the slides with collagen, laminin or fibronectin.

#3 bguyonnet

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Posted 23 September 2010 - 01:51 PM

Hi,

Having trouble in staining sperm cells that I'm taking out of the epididydmis with anti ubiquitin antibody.

for some reason the cells fell off the slides. I did methanol fixation for 15 min. and the antibody didn't stain at all although the data sheet say it is good for staining.

does any one have any tips??

something???

please

I going carzy.

thanks

tola


I am working on sperm cells also. For my IIF, I spread spermatozoa on Superfrost Excell slides from Fisher. I let them air-dried and do a methanol permeabilization and that works great!




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