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Antibody raised in mouse used on mouse tissue?

Antibody raised in mouse used on mouse tissue? - Western Blot Forum

Antibody raised in mouse used on mouse tissue? - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 11-06-2009, 08:39 PM
Pipette Filler
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Default Antibody raised in mouse used on mouse tissue?



For immunohisto there are several products that can help you use antibodies raised in mice on mouse tissues.

Is there anything like that for Western Blot analysis? There is only the one good antibody for my protein and therefore I am stuck using an antibody raised in mice on mouse tissues.

Special blocking agents or anything?

I am going to try to keep reducing the antibody concentration but there is only so low I can go.
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Old 04-16-2010, 01:46 PM
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Default Re: Antibody raised in mouse used on mouse tissue?

Hi,
using mouse antibodies in mouse samples has the problem that endogenous IgGs are stain together with your own antibody. However, if the MW of your protein is different that the MW for heavy (~50kDa) and light (~20kDa) chain of IgGs, you should not have any problem to see it.
I haven't tried but I suppose you can try to block the endogenous IgGs with for example goat anti-mouse IgG (without any further conjugation) before put your first antibody, but it could be a bit expensive depending in theconcentration of the blocking buffer.
In other hand, you have to consider how good is your antibody. Have you tried the westernblot with proteins from another specie? How many bands do you get in this case?
Regards,
Ileana
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