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Author Topic: ABC method conc?  (Read 3609 times)

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Offline Jason

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ABC method conc?
« on: August 25, 2004, 12:25:42 PM »
Hello,
 For doing the ABC method, what are the concentrations of the Avidin and the Biotin-HRP?
-Jason

If you can answer this 9:45 AM PST 08/25/04
it would be appreciated

ABC method conc?
« on: August 25, 2004, 12:25:42 PM »

Offline Jossy

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ABC method conc?
« Reply #1 on: August 25, 2004, 07:20:30 PM »
For IHC (in brain sections) I used the ABC kit (from Dako) 1:110, the Vectastain Kit also work 1:100 (aprox). remember that must be mixed 30 minutes before use (because the Avidin- biotin complex must be formed when use it)

Good Luck :D
ossy
jossy@neurobio.pitt.edu

Offline immunoem

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ABC method conc?
« Reply #2 on: August 26, 2004, 03:18:35 AM »
We use VectorLabs ABC kits (peroxidase and AP). For wax sections we use the A and B reagents each at 1:200.  For free-floating vibratome sections, where the incubation times are longer, we use each at 1:1000. As Jason mentions, the solution should be made 30 min in advance.
Good luck.
Ian

Offline immunoem

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ABC method conc?
« Reply #3 on: August 26, 2004, 03:20:51 AM »
Oops, should have said 'Jossy' not 'Jason'. Sorry 'bout that, haven't woken up fully yet - need coffee!
Ian

Offline richard03

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ABC method conc?
« Reply #4 on: August 26, 2004, 09:17:26 AM »
I used to use ABC kit from Vector Labs.  I have switched to HRP-Streptavidin which is also from Vector Labs.  I use 1:500 dilution in PBS, pH 7.2. The HRP-Streptavidin is very convenient since you don't need to wait for 30 minutes before incubation. I have been using this for many years and already completely switched to HRP-Streptavidin Solution. Accourding to a paper (I can't find the paper anymore), HRP-Streptavidin is about 5 times more sensitive than Standard ABC kit.

Hope I can find the paper and I will post it here.

Richard

Offline Jossy

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ABC method conc?
« Reply #5 on: August 26, 2004, 11:03:54 AM »
Hi, I always use it "as manufacturer recomends", but is good to know that i can use it more diluted   :D ......."learning never finish"
Thanks
ossy
jossy@neurobio.pitt.edu

ABC method conc?
« Reply #5 on: August 26, 2004, 11:03:54 AM »