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PBS in fractions

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Old 04-08-2008, 04:57 PM
Pipette Filler
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Default PBS in fractions
Hi i have a problem whit analyzing fractions which contain phosphate (20 mM) on WB i use BCIP/NBT substre for my secondary antibodies and there are no bands where they shuld be. Dose anybody know if phosphate ist the problem?

Thanks
Jasmina
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  #2 (permalink)  
Old 04-08-2008, 05:49 PM
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Default Re: PBS in fractions
PBS interferes with Alkaline phosphatase according to this source --> http://www.univ.trieste.it/~sc_biolo...20Blotting.pdf

I wasn't aware of it either.
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Old 04-09-2008, 02:59 AM
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Default Re: PBS in fractions
hmm never even knew that

thanks

I guess you will have to do a dialysis several times.
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Old 05-13-2008, 10:30 AM
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Default Re: PBS in fractions
hi,

try usinf TBS instead of PBS. i got sometimes also some high background using PBS and with TBS i got better results




greetz
alex
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