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Author Topic: Antigen Retrieval for PDGFR(alpha) rabbit  (Read 6376 times)

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Offline aztounded1

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Antigen Retrieval for PDGFR(alpha) rabbit
« on: February 09, 2006, 03:47:55 AM »
Can you A/R too far, not to where you have over staining, the staining image is like a pale gray, like a ghost image of where the antibody should be staining.

Antigen Retrieval for PDGFR(alpha) rabbit
« on: February 09, 2006, 03:47:55 AM »

Offline richard03

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Antigen Retrieval for PDGFR(alpha) rabbit
« Reply #1 on: February 11, 2006, 11:32:23 AM »
Can you post images? What substrate did you use to get the gray color, DAB?

Offline aztounded1

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Antigen Retrieval for PDGFR(alpha) rabbit
« Reply #2 on: February 16, 2006, 10:11:05 AM »
I used DAB+ from Dako for 5 minutes, I can send an image later today, from work, I also A/R the slide for 1 minute at 125C in Citrate (6.0), it's a rabbit antibody at a dilution of 1/100 in a PBS diluent.

Thanks.

Offline richard03

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Antigen Retrieval for PDGFR(alpha) rabbit
« Reply #3 on: February 27, 2006, 10:51:00 PM »
There are several possibilities:

1) DAB expired
2) AR too short, normally should be ~20 minutes
3) Tissue may be under-fixed.

Offline aztounded1

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Antigen Retrieval for PDGFR(alpha) rabbit
« Reply #4 on: February 28, 2006, 03:55:12 AM »
I have tried to A/R it longer, but I get everything staining, when I backed off the A/R and temp the stain finally worked, I used a different diluent and wow the stain works great, they say that the diluent is everything, I am starting to believe that saying.
   Thank you for the great advise, I did check all the things that were mentioned, but I never tried trying changing diluents.

Aztounded1

Antigen Retrieval for PDGFR(alpha) rabbit
« Reply #4 on: February 28, 2006, 03:55:12 AM »