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Post subject: IgG primary and IgM secondary
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Hello All,
I have a pretty basic question regarding immuno -
Is it possible to use an IgM biotinylated secondary antibody to detect an IgG primary antibody?
Any info is greatly appreciated,
Regards,
Science201! |
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 Sun Sep 18, 2011 3:35 am |
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Post subject: Welcome, Science 201
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Not quite sure what you mean...
If your IgM-biotin secondary is anti IgG, no problemo, if I understand correctly.
NB: I sometimes use IgG anti IgG-biotin secondary to detect IgM primaries ( I forget to use anti IgM.... .
I compared it with IgG -biotin that is anti IgM and could not see any differences.
Carl |
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| Sun Sep 18, 2011 4:47 pm |
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Post subject:
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Thanks Carl,
In fact I have done the opposite of what you sometimes do. I have used an anti IgM antibody to detect an IgG primary. I did not get any staining, using a positive control also, and I am hoping that the reason is that I used the incorrect secondary?
Thanks again! |
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| Sun Sep 18, 2011 10:54 pm |
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Post subject:
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Thanks Carl,
In fact I have done the opposite of what you sometimes do. I have used an anti IgM antibody to detect an IgG primary. I did not get any staining, using a positive control also, and I am hoping that the reason is that I used the incorrect secondary?
Thanks again! |
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| Sun Sep 18, 2011 10:54 pm |
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Post subject:
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Thanks Carl,
In fact I have done the opposite of what you sometimes do. I have used an anti IgM antibody to detect an IgG primary. I did not get any staining, using a positive control also, and I am hoping that the reason is that I used the incorrect secondary?
Thanks again! |
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| Sun Sep 18, 2011 10:55 pm |
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Post subject: Thanks for more info...
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If you have included a positive control ( an IgG primary) and that was also negative, you must be right in your assumption......I do not know why.
I hope others will inform us.
Good luck...
Carl |
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| Tue Sep 20, 2011 4:22 pm |
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Post subject: Re: Thanks for more info...
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| Carl wrote: |
If you have included a positive control ( an IgG primary) and that was also negative, you must be right in your assumption......I do not know why.
I hope others will inform us.
Good luck...
Carl |
Obviously Carl´s anti-IgG is directed to an Fc epitope common for IgG and IgM whereas Science201´s anti-IgM antibody is specific for IgM (probably an epitope in the core of the IgM antibody). |
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| Tue Oct 18, 2011 6:52 am |
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