Isotype Question - (Mar/24/2007 )

Quick question about isotypes....

I am wondeing if the specification sheet for my primary antibody gives a specific isotype for the immunogen (such as IgG2b species mouse) should I use a secondary specific for that isotype or will something general like the pierce anti-mouse HRP secondary work? This is for western blotting.

Also, can I save and reuse MOPS running buffer or do I have to make it up fresh each time? Can I save and reuse transfer buffer?

Thanks for your help.

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It does'nt matter. You could use secondary Ab different isotypes because of sec Ab specific for conservative Fc fragment of first Ab. For WB isotype is not important.
About tranfer buffer - keep at +4 and use one more foe the next WB ( if you do it during a week). For the time between WB more than 1 week we prepare fresh buffer. if you worry about MeOH use lower grade reagent for WB. You can use EtOH for WB but MEOH is preferable

"Also, can I save and reuse MOPS running buffer or do I have to make it up fresh each time?"

Before running, the gel doesn't have any etbr in it. This allows you to re-use the 1x MOPS running buffer several times

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