Re: Lectin staining

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From:"Barry Rittman" <brittman@mail.db.uth.tmc.edu>
To:histology <histonet@pathology.swmed.edu>
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Andrea
have never used tomato lectin but have done lot of lectin binding.
Lectin binding is very specific and many of the principles that apply to IHC
are also applicable to these.
The main problems is to ensure that you carry out a titre for the specific
lectin sample you have using a wide range of concentrations.
Don't know whether you are using a lectin that is conjugated or not. If
conjugated, I have found that some peroxidase conjugated lectins tend to brek
down and liberate peroxidase when stored at 4C.
Essential to use an affinity purified lectin and a good set of controls.
With certain lectins, the  pH and the presence of various ions such as
calcium and magnesium are also important.,
Tomato lectin binds to N-acetylglucosamine subunits. Could you let us know if
there are specific structures that you wish to demonstrate with this lectin.
As far as I remember glycophorin is a fairly good inhibitor.
thanks
Please contact me if you would like come more help with this.
Barry

AndreaH@imclone.com wrote:

> Does anyone do tomato lectin (or other lectin) staining? If so, could you
> share your protocol with me? Thank you in advance.
>
> Andrea
> andreah@imclone.com




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