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Author Topic: give me an anwer for antigen retrieval  (Read 5071 times)

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Offline daiyan1828

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give me an anwer for antigen retrieval
« on: September 08, 2005, 11:05:41 PM »
at first ,I come from China.nangjing Drum tower hospital.
I find a question on antigen require,the following
"Tris-EDTA pH 9:
Tris (Sigma T-1378) 1,211 g
EDTA "Tritriplex III (Merck 8418) 0, 372 g
Destilled water 1000 ml.

No pH ajustment is needed."
question :
1why  are pH 9?how to get pH 8
2why are Tris (Sigma T-1378) 1,211 g .in my place ,it is121g.
3why not adjust ?
4I find it do well in nucleolus .Can you agree it?
thanks

give me an anwer for antigen retrieval
« on: September 08, 2005, 11:05:41 PM »

Offline frank efterlon

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Re: give me an anwer for antigen require
« Reply #1 on: September 09, 2005, 06:07:28 AM »
Hi,

1 Adjust the pH with HCl.
2 It depends on the concentration you want. 121 g in 1 l is a 1M concentration. 1,21 g in 1 l is 0,01 M. In IHC 0,01 M is common use.
3 You can always adjust with HCl.

Frank

Offline madeleine huey

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give me an anwer for antigen retrieval
« Reply #2 on: September 12, 2005, 05:29:55 PM »
Hello,
EDTA come in 2 forms in dry chemicals.  It could be base or acid, didn't matter which one you use (use what's availuable in your lab).  The only thing you have to remember is adjust the ph to your need for EDTA.   For instance, most target retrieval for nuclear staining work better with EDTA, ph 9.0.  But that could be adjust to whatever work for your antigen in your tissue (but start with 9.0 first).  If you need ph = 6.0, then use Citrate Buffer.  Rather than EDTA, ph 9.  My experiences with these 2 buffer is, Citrate tend to give you a cleaner background, but intensity is weaker than EDTA.  For instance, if you get staining with Citrate, 6.0.  Then there's no need to go to EDTA, 9 (High alkaline with a ph of 9 tend to cause tissue falling off the slides, especially with microwave, but not so much with pressure cooker).  Hope this inf. give you the answer.
Madeleine

give me an anwer for antigen retrieval
« Reply #2 on: September 12, 2005, 05:29:55 PM »