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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
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1. BFP + 2. |
BCR - AB + | BCR + AB + | Overlay |
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![]() | ![]() |
Image Added
Discussion:
title | Plate map |
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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 100ngExpand | |||||||||||||||
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Expand | ||
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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. |
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hEF1a:eBFP hEF1a:GMabH hEF1a:GmabL hEF1a:CD79A hEF1a:CD79B |
hEF1a:CD79A |
hEF1a:CD79B |
aBeta 500 nM
Strep 10 ug/mL
![](/confluence/download/thumbnails/110860120/Screen%20Shot%202015-07-16%20at%2010.38.11%20PM.png?version=1&modificationDate=1437100983000&api=v2)
![](/confluence/download/thumbnails/110860120/Screen%20Shot%202015-07-16%20at%2010.38.48%20PM.png?version=1&modificationDate=1437101001000&api=v2)
![](/confluence/download/thumbnails/110860120/Screen%20Shot%202015-07-16%20at%2010.39.15%20PM.png?version=1&modificationDate=1437101025000&api=v2)
![](/confluence/download/thumbnails/110860120/Screen%20Shot%202015-07-16%20at%2010.39.55%20PM.png?version=1&modificationDate=1437101037000&api=v2)
![](/confluence/download/thumbnails/110860120/Screen%20Shot%202015-07-16%20at%2010.40.20%20PM.png?version=1&modificationDate=1437101054000&api=v2)
![](/confluence/download/thumbnails/110860120/Screen%20Shot%202015-07-16%20at%2010.46.27%20PM.png?version=2&modificationDate=1437101331000&api=v2)
![](/confluence/download/thumbnails/110860120/Screen%20Shot%202015-07-16%20at%2010.46.47%20PM.png?version=1&modificationDate=1437101370000&api=v2)
![](/confluence/download/thumbnails/110860120/Screen%20Shot%202015-07-16%20at%2010.47.26%20PM.png?version=1&modificationDate=1437101404000&api=v2)
![](/confluence/download/thumbnails/110860120/Screen%20Shot%202015-07-16%20at%2010.47.08%20PM.png?version=1&modificationDate=1437101390000&api=v2)
![](/confluence/download/thumbnails/110860120/Screen%20Shot%202015-07-16%20at%2010.47.39%20PM.png?version=1&modificationDate=1437101455000&api=v2)
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 +
Image Removed
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