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If I remember correctly acetylated BSA is supposed to counteract this binding to +ve charges as well as acting as a blocking agent. We find it works well with plant material. No need for a separate blocking step just put it in with the antibody. Has anyone used this approach with animal cells ?


Dr Lloyd Donaldson
Project Leader - Microscopy & Wood Identification
Senior Scientist - Plant Cell Walls & Biomaterials
Scion - Forests, Products, Innovation
49 Sala Street, Rotorua 3010
New Zealand
Ph 07 343 5581
www.scionresearch.com




-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of [log in to unmask]
Sent: Friday, 22 March 2013 10:39 a.m.
To: [log in to unmask]
Subject: Re: Blocking in immunocytochemsitry - ImageIT FX Enhancer?

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Not sure if this will apply to animal tissues, but for a couple of plant tissues where we kept getting background we blocked with pectin (started with pectin for jam - too many other ingredients, but several purified pectins are very cheap), and have also blocked with starch in the past (it was instant mashed potatoes). But the comments about miscellaneous binding to positive charges do resonate & will think about this in future.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E [log in to unmask]


On 21/03/13 8:23 PM, "Christophe Leterrier"
<[log in to unmask]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Hi,
>
>Not a confocal question, but a lot of people are doing immuno here so I
>thought I'd ask. We are currently using gelatin as the main blocker in
>our imunocytochemistry protocol. I was wondering if we could get better
>staining (less background / less non specific labeling) by switching to
>something else and found Life Technologie's ImageIT FX Enhancer (
>http://products.invitrogen.com/ivgn/product/I36933). Does someone have
>experience with this, and is the added cost justified? I try to stay
>clear of proprietary formulas but sometimes (see Prolong Gold) a "secret"
>product
>does work better.
>
>Thanks for the advices,
>
>Christophe
>
>--
>Christophe Leterrier
>Researcher
>Axonal Domains Architecture Team
>CRN2M CNRS UMR 7286
>Aix Marseille University, France



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