Fluoro-Immune Mice

So, antibodies are in use in many many many labs, right here in our lab, there are over 1000 different types. They're incredibly useful. In ideal conditions, they can be used to detect a single variant of a single protein among 1000s. Which is nice.

Now, I want to improve them. Simply genetically modify mice/rats/rabbits to produce all of their antibodies with a fluorescent protein tag, such as CFP/YFP/GFP etc.

Then, simply produce antibodies in the normal way: Inoculate, wait a bit, harvest & purify.

There are several reasons why this is great:

1. Their immune system will be fluorescent, which is fun, but not immediately useful to non-immunologists.

2. Antibody purification and quantification may be more effective when assisted and monitored by fluorometry.

3. They will not require secondary antibodies for Immunofluorescence and fluorescent Western Blot experiments. This a: saves between 30mins and 1hr for every experiment of this type ever performed. b: removes 1 layer of complexity and source of false positives.

4. The resolution of microscopy experiments will be improved: Normally, the fluorophore is on the end of the secondary antibody, which is attached to the primary antibody. Each antibody is about 10nm, so, the fluorophore could be anywhere in a 20nm radius of the protein of interest. This cuts that to 10nm. So you can get a better idea of where things are, and it's useful for super- resolution microscopy<link>, which is becoming all easy- peasy lemon-squeezy

5. Co-localization experiments are enhanced by the possibility of using fluorescence resonance electron transfer (FRET)<link>. Normally, you can use 2 separate antibodies to probe for two independent proteins, and look to see if they're in the same place, sort of. Now, it's very difficult to draw strong conclusions from these data, as a microscope's resolution is a few hundred nm. Many proteins that really don't interact at all can be found within a few hundred nm... so it's vague supportive evidence at best. Looks good though, hence it's persistence in the literature. Now, FRET, only works if the proteins are REALLY close, so the CFP antibody you got from a mouse and the YFP antibody from a Rabbit only produce a positive co-localization in a FRET experiment if they're right next to each other or directly interacting.

6. You can attach some of those whizz-bang photoswitchable proteins, like Dronpa, to antibodies... 'cause, you know they're what the with-it kids are all about these days.

Right, got to go and wash my secondary antibody off....