Author Topic: Re: Welcome!  (Read 2697 times)

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Offline stathis

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Re: Welcome!
« on: March 15, 2003, 11:33:27 AM »
Hi everyone,
Does the use of 1% Triton X-100 in the wash solutions for IHC
may cause problems concerning with non-specific binding?

STHTHIS

Re: Welcome!
« on: March 15, 2003, 11:33:27 AM »

Offline susan

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1% Triton X-100
« Reply #1 on: March 15, 2003, 12:35:53 PM »
Triton X-100 is a very good detergent for IHC to enhance the penetration of antibodies. The appropriate use of Triton X-100 will produce superior results. Triton X-100 is normally used in blocking buffer and dilution buffer and rarely used in washing solution, which may cause no-specific binding.

The concentration of Triton X-100 is also critical. 1% Triton X-100 is too high for IHC purpose. Normally, the concentration of 0.2% is good for LM, and 0.02%-0.04% is good for EM.

Hope this helps.

Susan

Offline wangjing_gene

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Re: 1% Triton X-100
« Reply #2 on: October 12, 2003, 12:13:36 AM »
Thanks Susan, it's helpful.  but I still want to confirm whether Triton x-100 is needed in the primary or 2nd AB staining solutions.

Quote from: "susan"
Triton X-100 is a very good detergent for IHC to enhance the penetration of antibodies. The appropriate use of Triton X-100 will produce superior results. Triton X-100 is normally used in blocking buffer and dilution buffer and rarely used in washing solution, which may cause no-specific binding.

The concentration of Triton X-100 is also critical. 1% Triton X-100 is too high for IHC purpose. Normally, the concentration of 0.2% is good for LM, and 0.02%-0.04% is good for EM.

Hope this helps.

Susan

Offline susan

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Triton X-100
« Reply #3 on: October 12, 2003, 09:32:33 AM »
For sections less than 10um thick, Triton X-100 is not needed in primary and 2nd dilution buffer. I do if sections are thicker such as 30~50 um to help Ab penetration.

Susan

Offline wangjing_gene

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Triton X-100
« Reply #4 on: October 12, 2003, 09:42:11 AM »
I c, thanks a lot!

Triton X-100
« Reply #4 on: October 12, 2003, 09:42:11 AM »