Background:
The literature is unclear on whether Lyn is always associated with the BCR complex or if it binds only upon BCR dimerization. We know from the literature that Syk will bind but only in the presence of Lyn or two other Src family kinases that are known to associate with the BCR. Lyn is known to be expressed by HEK 293 cells so there may be competition with the binding of our engineered versions of syk-TEV and lyn-TEV. The purpose of this experiment is to determine whether or not Lyn will function as a cofactor in the TANGO system with the BCR. Essentially we want to know if Lyn is always associated with the BCR or if it is recruited upon activation of the receptor. We will be using Gmab because we have not yet ordered or constructed any other antibodies.
Parts Needed:
- Gmab H
- Gmab L
- Syk-TEV
- Lyn-TEV
- Lyn
- CD79A
- CD79B
- CD79A-TCS-G4
- CD79B-TCS-G4
- UAS: mKate
- hEF1a: BFP
- abeta
- hEF1a: G4
Procedure:
| HEK | Positive Control | Negative Control | |||||||||
| Syk => | |||||||||||
| Lyn => | |||||||||||
Results:
Discussion: