How to detect specific-protein producing cells? - (Jan/06/2016 )
Hi
I purchased Streptomyces Avidinii from DSMZ for the aim of Streptavidin production. I know from literature that many of DSMZ's bacteria my lost their ability to produce Streptavidin and my first task is detect cells that is producing.
Use a labeled biotin probe and a FACS.
bob1 on Mon Jan 11 20:12:38 2016 said:
Use a labeled biotin probe and a FACS.
thank you, but I dont have fluorescent equipments and I look for a simple and cheap assay.
OK, I was under the assumption that you were looking to isolate the individual cells. For a cell population you could do this with an ELISA for streptavidin/biotin binding
bob1 on Sun Jan 17 07:56:38 2016 said:
OK, I was under the assumption that you were looking to isolate the individual cells. For a cell population you could do this with an ELISA for streptavidin/biotin binding
can you explain it a little more?
am I right? you mean we coat our supernatant (that contained streptavidin or not) in plate and then add for example biotinylated HRP and then add indicator reagent
Arash Rostami on Sun Jan 17 12:59:21 2016 said:
bob1 on Sun Jan 17 07:56:38 2016 said:
OK, I was under the assumption that you were looking to isolate the individual cells. For a cell population you could do this with an ELISA for streptavidin/biotin binding
can you explain it a little more?
am I right? you mean we coat our supernatant (that contained streptavidin or not) in plate and then add for example biotinylated HRP and then add indicator reagent
Yes, that's the general idea. You might have to play around with conditions a bit, but there should be good protocols on the web.
bob1 on Sun Jan 17 20:54:38 2016 said:
Arash Rostami on Sun Jan 17 12:59:21 2016 said:
bob1 on Sun Jan 17 07:56:38 2016 said:
OK, I was under the assumption that you were looking to isolate the individual cells. For a cell population you could do this with an ELISA for streptavidin/biotin binding
can you explain it a little more?
am I right? you mean we coat our supernatant (that contained streptavidin or not) in plate and then add for example biotinylated HRP and then add indicator reagent
Yes, that's the general idea. You might have to play around with conditions a bit, but there should be good protocols on the web.
Thank you so much
I am confused, for the elisa-like procedure can I first coat supernatant of bacteria and then add biotinylated HRP and see change of color? does it work?
I cant use antibody it's expensive .
Should be fine.