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How to detect specific-protein producing cells?


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#1 Arash Rostami

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Posted 06 January 2016 - 06:45 AM

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.

 
I wonder if you help me,please.


#2 bob1

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Posted 11 January 2016 - 12:12 PM

Use a labeled biotin probe and a FACS.



#3 Arash Rostami

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Posted 16 January 2016 - 12:55 PM

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. 



#4 bob1

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Posted 16 January 2016 - 11:56 PM

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



#5 Arash Rostami

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Posted 17 January 2016 - 04:59 AM

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


Edited by Arash Rostami, 17 January 2016 - 05:01 AM.


#6 bob1

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Posted 17 January 2016 - 12:54 PM

 

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.



#7 Arash Rostami

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Posted 19 January 2016 - 04:17 PM

 

 

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 muchsmile.png



#8 Arash Rostami

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Posted 06 February 2016 - 11:12 PM

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 .



#9 bob1

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Posted 07 February 2016 - 01:04 AM

Should be fine.






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