Review of blocking reagants.

[Kevin Sunley]

Hello everyone,

While this question is specifically confocal-related, I was wondering if
anyone knew a good, comprehensive, review of different blocking reagents
(commercially available, or homebrew) for immunofluorescence staining.

Aside from just listing the standards; x% BSA or milk powder, different
percents of serum, and the commercially available options (Image IT,
etc.), has anyone ever seen a paper that reviews the effectiveness of
these in relation to each other.

Alternatively, if its not in a paper format, has anyone compared these
themselves and found a favourite?


Thanks in advance,


Kevin Sunley
University of Manitoba
Winnipeg, Canada

[George McNamara]

Hi Kevin,

Check out:


Immunofluorescence imaging of DNA damage response proteins: optimizing protocols for super-resolution microscopy.
Bennett BT, Bewersdorf J, Knight KL.
Methods. 2009 May;48(1):63-71. Epub 2009 Feb 24.
PMID: 19245833

as well as Invitrogen's web site for Image-iT FX.

May all your staining be specific,

George



At 08:25 AM 6/4/2009, you wrote:

Hello everyone,

While this question is specifically confocal-related, I was wondering if anyone knew a good, comprehensive, review of different blocking reagents (commercially available, or homebrew) for immunofluorescence staining.

Aside from just listing the standards; x% BSA or milk powder, different percents of serum, and the commercially available options (Image IT, etc.), has anyone ever seen a paper that reviews the effectiveness of these in relation to each other.

Alternatively, if its not in a paper format, has anyone compared these themselves and found a favourite?


Thanks in advance,


Kevin Sunley
University of Manitoba
Winnipeg, Canada

[Mayandi Sivaguru]

Kevin, In my experience for quick preps with goat secondary antibodies the Invitrogen Image IT works great. However, for economical and yet effective, for all other routine preps I recommend Tom's protocol as in this link works great and comes for long time.

http://core.igb.uiuc.edu/Blocking%20Protocol%20LM_EM.pdf
Shiv

At 07:25 AM 6/4/2009, you wrote:

Hello everyone,

While this question is specifically confocal-related, I was wondering if anyone knew a good, comprehensive, review of different blocking reagents (commercially available, or homebrew) for immunofluorescence staining.

Aside from just listing the standards; x% BSA or milk powder, different percents of serum, and the commercially available options (Image IT, etc.), has anyone ever seen a paper that reviews the effectiveness of these in relation to each other.

Alternatively, if its not in a paper format, has anyone compared these themselves and found a favourite?


Thanks in advance,


Kevin Sunley
University of Manitoba
Winnipeg, Canada

Mayandi Sivaguru, PhD, PhD
Microscopy Facility Manager
8, Institute for Genomic Biology
University of Illinois at Urbana-Champaign
1206 West Gregory Dr.
Urbana, IL 61801 USA

Office: 217.333.1214
Fax: [phone]
[hidden email]
http://core.igb.uiuc.edu