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Offline WC

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Controls and specificty of IHC tests
« on: April 28, 2009, 11:58:07 PM »
Can someone tell a non scientist doing some related research whether the following are specific to (deep) nerve tissue (eg brain/spinal cord);  GFAP, NF, S100, SYN.  

Also, are there any agreed industry protocols for IHC when it comes to controls? What for example are the minimum requirements for positive controls?  Hope these questions aren't too imprecise!  Thanks in advance.

Controls and specificty of IHC tests
« on: April 28, 2009, 11:58:07 PM »

Offline tony1

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Re: Controls and specificty of IHC tests
« Reply #1 on: June 07, 2009, 03:54:28 PM »
GFAP will pick out the glial cells and NFP will pick out the neurons, these should be demonstrated in any tissue of neural origin.

S100 will also demonstrate the nerves and is pretty good at it.

Positive controls should be used for each batch of slides of a particular antibody, e.g. if doing 30 S100s then a minimum of 1 S100 control, although experience has shown me that the best thing to do if you have enough control tissue is to put the test and control on the same slide, reduces the number of controls, reduces the amount of primary antisera and is the only real guarantee that the control has had the same treatment as the test.

The most important thing to consider is the negative control, there should be a minimum of one per different type of primary antisera. The replacement for the primary antisera can be the antibody diluent, but is better being a non immune serum to the same concentration of the primary antibody at the working dilution.

Hope this helps

Tony Crick
www.hesyre.com
The health professional community

Offline tony1

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Re: Controls and specificty of IHC tests
« Reply #2 on: June 07, 2009, 03:56:41 PM »
oops forgot to mention may be worth doing double staining?

Re: Controls and specificty of IHC tests
« Reply #2 on: June 07, 2009, 03:56:41 PM »