| Expand | ||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| ||||||||||||||||||
|
Results:
Untransfected Control | Single Color Control | Background Beta Amyloid Binding | Background Strep Binding | Beta Amyloid Binding to Gantenerumab |
Well 1HEK293 | Well 2HEK293 hEF1a:eBFP 100ng Dummy 400ng | Well 3HEK293 hEF1a:eBFP 100ng Dummy 400ng aBeta 500 nM Strep 10 ug/mL
| Well 4HEK293 hEF1a:eBFP 100ng hEF1a:GMabH 100ng hEF1a:GMabL 100ng hEF1a:CD79A 100ng hEF1a:CD79B 100ng Strep 10 ug/mL | Well 5HEK293 hEF1a:eBFP 100ng hEF1a:GMabH 100ng hEF1a:GMabL 100ng hEF1a:CD79A 100ng hEF1a:CD79B 100ng aBeta 500 nM Strep 10 ug/mL
|
 |  |  |  |  |
 |  |  |  |  |
| Expand | ||
|---|---|---|
| ||
1. BFP + 2.
|
BCR - AB + | BCR + AB + | Overlay |
|---|---|---|
 |  |
Discussion:
| title | Plate map |
|---|
Untransfected
Control
Single Color
Control
Background Beta
Amyloid Binding
Background
Strep Binding
Beta Amyloid Binding
to Gantenerumab
Well 1
HEK293
Well 2
HEK293
hEF1a:eBFP 100ng
Dummy 400ng
Well 3
HEK293
hEF1a:eBFP 100ng
Dummy 400ng
aBeta 500 nM
Strep 10 ug/mL
Well 4
HEK293
hEF1a:eBFP 100ng
hEF1a:GMabH 100ng
hEF1a:GMabL 100ng
hEF1a:CD79A 100ng
hEF1a:CD79B 100ng
Strep 10 ug/mL
Well 5
HEK293
hEF1a:eBFP 100ng
hEF1a:GMabH 100ng
hEF1a:GMabL 100ng
hEF1a:CD79A 100ng
hEF1a:CD79B 100ng
aBeta 500 nM
Strep 10 ug/mL
Well 7
HEK293
Well 8
HEK293
hEF1a:eBFP 100ng
Dummy 400ng
Well 9
HEK293
hEF1a:eBFP 100ng
Dummy 400ng
aBeta 500 nM
Strep 10 ug/mL
Well 10
HEK293
hEF1a:eBFP 100ng
hEF1a:GMabH 100ng
hEF1a:GMabL 100ng
hEF1a:CD79A 100ng
hEF1a:CD79B 100ng
Strep 10 ug/mL
Well 11
HEK293
hEF1a:eBFP 100ng
hEF1a:GMabH 100ng
hEF1a:GMabL 100ng
hEF1a:CD79A 100ng
hEF1a:CD79B 100ng
aBeta 500 nM
Strep 10 ug/mL
Results:
Untransfected
Control
Single Color
Control
Background Beta
Amyloid Binding
Background
Strep Binding
Beta Amyloid Binding
to Gantenerumab
Well 1
HEK293
Well 2
HEK293
hEF1a:eBFP 100ng
Dummy 400ng
Well 3
HEK293
hEF1a:eBFP 100ng
Dummy 400ng
aBeta 500 nM
Strep 10 ug/mL
Well 4
HEK293
hEF1a:eBFP 100ng
hEF1a:GMabH 100ng
hEF1a:GMabL 100ng
hEF1a:CD79A 100ng
hEF1a:CD79B 100ng
Strep 10 ug/mL
Well 5
HEK293
hEF1a:eBFP 100ng
hEF1a:GMabH 100ng
hEF1a:GMabL 100ng| Expand | |||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| |||||||||||||||
|
| Expand | ||
|---|---|---|
| ||
Testing Gantenerumab binding to beta amyloid oligomers in the context of the B-Cell Receptor. Repeat of experiment done during iGEM. |
| Expand | ||
|---|---|---|
| ||
Testing beta amyloid binding to cells expressing the components of the B-Cell Receptor. To do this, we are going to treat cells with a saturating concentration of biotin-conjugated beta amyloid. Subsequently, we are going to treat with AlexaFluor 594 conjugated streptavidin. If beta-amyloid binds to the receptor, streptavidin can bind biotin and be detected via flow cytometry. We are controlling for eBFP bleed through into the red channel, background beta-amyloid binding and background streptavidin binding. |
| Expand | ||
|---|---|---|
| ||
hEF1a:eBFP hEF1a:GMabH hEF1a:GmabL hEF1a:CD79A hEF1a:CD79B |
| title | Gating |
|---|
1. BFP +
2.
Untransfected
Control
Single Color
Control
Background Beta
Amyloid Binding
Background
Strep Binding
Beta Amyloid Binding
to Gantenerumab
Well 1
HEK293
Well 2
HEK293
hEF1a:eBFP 100ng
Dummy 400ngWell 3
HEK293
hEF1a:eBFP 100ng
Dummy 400ng
aBeta 500 nM
Strep 10 ug/mL
Well 4
HEK293
hEF1a:eBFP 100ng
hEF1a:GMabH 100ng
hEF1a:GMabL 100ng
hEF1a:CD79A 100ng
hEF1a:CD79B 100ng
Strep 10 ug/mL
Well 5
HEK293
hEF1a:eBFP 100ng
hEF1a:GMabH 100ng
hEF1a:GMabL 100ng
hEF1a:CD79A 100ng
hEF1a:CD79B 100ng
aBeta 500 nM
Strep 10 ug/mL
BCR -
AB +
AB +
Discussion:
| Expand | |||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| |||||||||||||||
|
| Expand | ||
|---|---|---|
| ||
Testing Gantenerumab binding to beta amyloid oligomers in the context of the B-Cell Receptor. Repeat of experiment done during iGEM. |
| Expand | ||
|---|---|---|
| ||
Testing beta amyloid binding to cells expressing the components of the B-Cell Receptor. To do this, we are going to treat cells with a saturating concentration of biotin-conjugated beta amyloid. Subsequently, we are going to treat with AlexaFluor 594 conjugated streptavidin. If beta-amyloid binds to the receptor, streptavidin can bind biotin and be detected via flow cytometry. We are controlling for eBFP bleed through into the red channel, background beta-amyloid binding and background streptavidin binding. |
| title | Parts Needed |
|---|
hEF1a:eBFP
hEF1a:GMabH
hEF1a:GmabL
hEF1a:CD79A
hEF1a:CD79B