Proteins needed(THIS IS OUT OF DATE) - Needed for primary experiments:
- Reporter circuit (activatable promoter + reporter Gal4UAS - (eYFP))
- Gal4UAS-mKate
- Tre-TEVp
- Hef1a-rtTa
- dox support circuit
- dox Syk+TEV measurement circuit (dox dependent same promoter as Syk+TEV(which is TRE) + mKate)
- CD79A
- normal
- fusion: CD79A C terminus + tev cut site + Transcriptional activatorGal4VP16
- CD79B
- normal
- fusion: CD79B C terminus + tev cut site + Transcriptional activatorGal4VP16
- Lyn
- normal
- fusion: FP eYFP 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))
- (TRE-Syk-TEVp) fusion, point mutation: 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 TCS +Transcriptional activatorGal4VP16
- Needed for debug experiments
we have eBFP(2?), eYFP, mKATE
Supplies needed - Dyed(dyeable?) anti IgM (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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