Proteins needed(THIS IS OUT OF DATE) - Needed for primary experiments:
- Reporter circuit (activatable promoter + reporter (FP))
- CD79A
- normal
- fusion: CD79A C terminus + tev cut site +
FP(mKATE)- Transcriptional activator
- CD79B
- normal
- fusion: CD79B C terminus + tev cut site +
FP(eBFP)- transcriptional activator
- Lyn
- normal
- fusion: FP tagged (N or C terminus?)
- point mutation: mutated to always phosphorylate
- Syk
- fusion, point mutation: FP attached(N or C terminus?), prevent phosphorylation (eYFP tagged SYK shouldn't be in same cell as FP anything else, ought be no cross talk)
- fusion, point mutatino: tev protease attached (N or C terminus?), point mutated to prevent phosphorylation
- ig k
- with proper localization tag
- igM heavy
- with proper constant region for membrane localization and proper localization tag
- dito + tev +eYFP
- Needed for debug experiments
we have eBFP(2?), eYFP, mKATE
Supplies needed - Antibody that targets membrane bound igM that is dyed so we can see it (we can buy this, probably target heavy chain, may also want light chain as well to make sure both localize)
- Beta amyloid (? some kind that binds to cells and some kind we can wash antibodies over?)
- Lyn and Syk phosphorylation assay
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