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Staining of antibodies...

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Benoit Labarthe Benoit Labarthe
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Staining of antibodies...

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear All,

I have to stain antibodies with an antibody staining kit designed for 100µg of protein from molecular probes, my problem is that I can not spend 100µg of my precious antibody for this staining... Do you think that staining of only 50 µg of antibody with this kit would be OK? Do you have any tips to improve the staining of low concentration/mass of antibodies with this type of staining kit?

Thank you

benoit



--
Benoît Labarthe, PharmD, MSc
Etudiant PhD - PhD trainee
Institut de Cardiologie de Montréal - Montreal Heart Institute
Centre de Recherche, Research Center
Laboratoire du Dr.Théroux
5000 Bélanger Est
Montréal Québec Canada H1T 1C8
Tel (514)-376 3330 ext 3017
Fax (514)- 376 1076
Scott Howell-3 Scott Howell-3
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Re: Staining of antibodies...

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Benoit,

If I am understanding you correctly it sounds like you might be better
off using a secondary antibody to "tag" your antibody instead of
directly tagging your primary. In this manner you will use a lot less.
Hope this helps.

Scott J. Howell, Ph.D.
Manager, Imaging Module
Visual Sciences Research Center
Case Western Reserve University
2085 Adelbert Rd.
Institute of Pathology Room 106
Cleveland, Ohio 44106
216-368-2300
http://www.case.edu/med/vsrc/

----- Original Message -----
From: Benoit Labarthe <[hidden email]>
Date: Tuesday, May 6, 2008 2:55 pm
Subject: Staining of antibodies...
To: [hidden email]

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear All,
>
> I have to stain antibodies with an antibody staining kit designed
> for 100µg
> of protein from molecular probes, my problem is that I can not
> spend 100µg
> of my precious antibody for this staining... Do you think that
> staining of
> only 50 µg of antibody with this kit would be OK? Do you have any
> tips to
> improve the staining of low concentration/mass of antibodies with
> this type
> of staining kit?
>
> Thank you
>
> benoit
>
>
>
> --
> Benoît Labarthe, PharmD, MSc
> Etudiant PhD - PhD trainee
> Institut de Cardiologie de Montréal - Montreal Heart Institute
> Centre de Recherche, Research Center
> Laboratoire du Dr.Théroux
> 5000 Bélanger Est
> Montréal Québec Canada H1T 1C8
> Tel (514)-376 3330 ext 3017
> Fax (514)- 376 1076
>
Ignatius, Mike Ignatius, Mike
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Re: Staining of antibodies...

In reply to this post by Benoit Labarthe
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
We designed those microscale labeling kits for 20 to 100ugs.  50 should be fine.
 
Mike Ignatius

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Benoit Labarthe
Sent: Tuesday, May 06, 2008 11:55 AM
To: [hidden email]
Subject: Staining of antibodies...

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear All,

I have to stain antibodies with an antibody staining kit designed for 100µg of protein from molecular probes, my problem is that I can not spend 100µg of my precious antibody for this staining... Do you think that staining of only 50 µg of antibody with this kit would be OK? Do you have any tips to improve the staining of low concentration/mass of antibodies with this type of staining kit?

Thank you

benoit



--
Benoît Labarthe, PharmD, MSc
Etudiant PhD - PhD trainee
Institut de Cardiologie de Montréal - Montreal Heart Institute
Centre de Recherche, Research Center
Laboratoire du Dr.Théroux
5000 Bélanger Est
Montréal Québec Canada H1T 1C8
Tel (514)-376 3330 ext 3017
Fax (514)- 376 1076
Benoit Labarthe Benoit Labarthe
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Re: Staining of antibodies...

In reply to this post by Scott Howell-3
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Thank you Scott

Sadly, we do not have this choice, our experiment does not allow us to use secondary stainings....

benoit




On Tue, May 6, 2008 at 3:02 PM, Scott Howell <[hidden email]> wrote:
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Benoit,

If I am understanding you correctly it sounds like you might be better
off using a secondary antibody to "tag" your antibody instead of
directly tagging your primary. In this manner you will use a lot less.
Hope this helps.

Scott J. Howell, Ph.D.
Manager, Imaging Module
Visual Sciences Research Center
Case Western Reserve University
2085 Adelbert Rd.
Institute of Pathology Room 106
Cleveland, Ohio 44106
216-368-2300
http://www.case.edu/med/vsrc/

----- Original Message -----
From: Benoit Labarthe <[hidden email]>
Date: Tuesday, May 6, 2008 2:55 pm
Subject: Staining of antibodies...
To: [hidden email]

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear All,
>
> I have to stain antibodies with an antibody staining kit designed
> for 100µg
> of protein from molecular probes, my problem is that I can not
> spend 100µg
> of my precious antibody for this staining... Do you think that
> staining of
> only 50 µg of antibody with this kit would be OK? Do you have any
> tips to
> improve the staining of low concentration/mass of antibodies with
> this type
> of staining kit?
>
> Thank you
>
> benoit
>
>
>
> --
> Benoît Labarthe, PharmD, MSc
> Etudiant PhD - PhD trainee
> Institut de Cardiologie de Montréal - Montreal Heart Institute
> Centre de Recherche, Research Center
> Laboratoire du Dr.Théroux
> 5000 Bélanger Est
> Montréal Québec Canada H1T 1C8
> Tel (514)-376 3330 ext 3017
> Fax (514)- 376 1076
>



--
Benoît Labarthe, PharmD, MSc
Etudiant PhD - PhD trainee
Institut de Cardiologie de Montréal - Montreal Heart Institute
Centre de Recherche, Research Center
Laboratoire du Dr.Théroux
5000 Bélanger Est
Montréal Québec Canada H1T 1C8
Tel (514)-376 3330 ext 3017
Fax (514)- 376 1076
Benoit Labarthe Benoit Labarthe
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Re: Staining of antibodies...

In reply to this post by Ignatius, Mike
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Good news! what are the critical points of the protocol, would an increase of the incubation time to 2h help for the staining?

benoit

On Tue, May 6, 2008 at 3:12 PM, Ignatius, Mike <[hidden email]> wrote:
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
We designed those microscale labeling kits for 20 to 100ugs.  50 should be fine.
 
Mike Ignatius

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Benoit Labarthe
Sent: Tuesday, May 06, 2008 11:55 AM

To: [hidden email]
Subject: Staining of antibodies...

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear All,


I have to stain antibodies with an antibody staining kit designed for 100µg of protein from molecular probes, my problem is that I can not spend 100µg of my precious antibody for this staining... Do you think that staining of only 50 µg of antibody with this kit would be OK? Do you have any tips to improve the staining of low concentration/mass of antibodies with this type of staining kit?

Thank you

benoit



--
Benoît Labarthe, PharmD, MSc
Etudiant PhD - PhD trainee
Institut de Cardiologie de Montréal - Montreal Heart Institute
Centre de Recherche, Research Center
Laboratoire du Dr.Théroux
5000 Bélanger Est
Montréal Québec Canada H1T 1C8
Tel (514)-376 3330 ext 3017
Fax (514)- 376 1076



--
Benoît Labarthe, PharmD, MSc
Etudiant PhD - PhD trainee
Institut de Cardiologie de Montréal - Montreal Heart Institute
Centre de Recherche, Research Center
Laboratoire du Dr.Théroux
5000 Bélanger Est
Montréal Québec Canada H1T 1C8
Tel (514)-376 3330 ext 3017
Fax (514)- 376 1076
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