Author Topic: pax7 and MyoD ICC  (Read 4739 times)

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Offline lemonpot

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pax7 and MyoD ICC
« on: January 19, 2011, 03:27:37 AM »
i am currently doing ICC for Pax7(mouse) and MyoD(rabbit) within a slide chamber.
My concentration for:
- Pax 7 primary Ab is 1:15 to 1:100 and secondary is alexa fluor 546(1:300)
- MyoD primary Ab is 1:15 to 1:100 as well,  secondary is donkey anti rabbit biotinylated(1:300) and tertiary is alexa fluor 488 strepavidin conjugated 488(1:400).
Blocker i am using is 5% goat & donkey serum in PBS with 0.35% carrageen.

the thing is MyoD signals comes out fine..however pax7 i can never get it to show..Neg control shows nothing...i have not tried positive controls.
any idea what is going wrong??

would appreciate is someone can help

pax7 and MyoD ICC
« on: January 19, 2011, 03:27:37 AM »

Offline MT Scientist

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Re: pax7 and MyoD ICC
« Reply #1 on: January 19, 2011, 11:49:33 AM »
Just an observation:
MyoD is done using a biotin amplification while your Pax7 is not.  It stands to reason, that if your Pax7 is also initially a faint signal, it should also be amplified....

Offline lemonpot

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Re: pax7 and MyoD ICC
« Reply #2 on: January 27, 2011, 10:01:14 PM »
I am kinda lost in the pax7 stainin...thinking of changing my Pax 7 antibody brand entirely

Re: pax7 and MyoD ICC
« Reply #2 on: January 27, 2011, 10:01:14 PM »