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Author Topic: TBS or PBS for washing?  (Read 21024 times)

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Offline arnotts

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TBS or PBS for washing?
« on: November 22, 2005, 01:56:05 AM »
Hi..
How do I know which is the best to use for washing? TBS or PBS?
And why does some protocols include Tween-20 in the buffer? what does it do?

Regards,
arnotts    :D

TBS or PBS for washing?
« on: November 22, 2005, 01:56:05 AM »

Offline wanglll

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Re: TBS or PBS for washing?
« Reply #1 on: November 22, 2005, 03:42:19 AM »
Generally, the TBS is better than PBS in the immunohistochemistry, but the cost of TBS is  expensive for PBS, the washing results of PBS is similitude with TBS in immunohistochemical stainers.

Tween 20 , also known generically as Polysorbate 20 is a surfactant and spreading agent, Tween 20 is useful in many immunohistochemical procedures and may be used on automated stainers which require a TBS or PBS wash. :shock:  :P

Offline jonnieplumb

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TBS or PBS for washing?
« Reply #2 on: November 23, 2005, 05:58:14 AM »
Hi I have found that depending on the tissue used and the detection method applied will govern whether I use PBS or TBS.
FFPE - TBS (diluent) with TBST wash
SF - PBS diluent and wash
IPx - PBS
AP - TBS and TBST

Offline ImmunoNYC

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TBS or PBS for washing?
« Reply #3 on: November 23, 2005, 11:13:58 PM »
I have always wanted to do specific buffers for specific stains but never had the time to devote to working this out.

So in lieu of this I work with what works for me ... I routinely use TBS with a little Tween as my diluent buffer for antibodies (including serum and BSA as needed etc). I use PBS for all manual stains and TBS/Tween for all autostainer stains.

Offline jkb

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TBS or PBS for washing?
« Reply #4 on: May 16, 2006, 04:00:12 PM »
Hi MaximinaNYC,
What percentage of BSA or serum do you add to your wash buffer for the antibody diluent?   I was planning on using TBST as a wash buffer and wasn't sure what the best option would be for the antibody diluent.  
Thank you.

Offline ImmunoNYC

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TBS or PBS for washing?
« Reply #5 on: May 16, 2006, 09:21:31 PM »
I use 1% BSA & 2% serum in TBS with 0.004% Tween-20 (just the tiniest amount) for antibody diluent. I have done this for years and I have absolutely NO background problems.

I do not add BSA or serum to wash buffers. The cost would be obscene.
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Offline jkb

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TBS or PBS for washing?
« Reply #6 on: May 17, 2006, 08:23:13 AM »
Thank you for your response.  Do you use serum from the host species of the secondary antibody?

Offline ImmunoNYC

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TBS or PBS for washing?
« Reply #7 on: May 17, 2006, 10:44:06 AM »
Yes.

Quote
Thank you for your response.  Do you use serum from the host species of the secondary antibody?

TBS or PBS for washing?
« Reply #7 on: May 17, 2006, 10:44:06 AM »