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titleSetup

 

 

Well 1

HEK293

Well 2

HEK293

Dummy DNA

eYFP

Streptavidin

Well 3

HEK293

Dummy DNA

eYFP

1000 nM Monomerized aBeta

Streptavidin

Well 4

HEK293

Dummy DNA

eYFP

1000 nM Oligomerized aBeta

Streptavidin

Well 5

HEK293

Gmab H

Gmab L

CD79A

CD79B

eYFP

Streptavidin

Well 6

HEK293

Gmab H

Gmab L

CD79A

CD79B

eYFP

1000 nM Monomerized aBeta

Streptavidin

Well 7

HEK293

Gmab H

Gmab L

CD79A

CD79B

eYFP

1000 nM Oligomerized aBeta

Streptavidin

Well 8

EMPTY

Well 9

EMPTY

Well 10

EMPTY

Well 11

EMPTY

Well 12

EMPTY

Well 13

HEK293

Dummy DNA

eYFP

StreptavidinEMPTY

Well 14EMPTY

HEK293

Dummy DNA

eYFP

Streptavidin

Beta-amyloid oligomers 500nM

Well 15

EMPTYHEK293

Well 16

HEK293

EMPTYStreptavidin

Well 17EMPTY

HEK293

Streptavidin

Beta-amyloid monomers 500nM

Well 18EMPTY

HEK293

Streptavidin

Beta-amyloid oligomers 500nM

Well 19

EMPTY

Well 20

EMPTY

Well 21

EMPTY

Well 22

EMPTY

Well 23

EMPTY

Well 24

EMPTY

 

 

 

Well 1

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

0 nM abeta oligomer 

Well 2

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

0.1 nM abeta oligomerStreptavidin

Well 3

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP0.25 nM abeta oligomer

Streptavidin

Beta-amyloid monomers 500nM

Well 4

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP0.5 nM abeta oligomer

Streptavidin

Beta-amyloid oligomers 500nM

Well 5

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

1 nM abeta oligomer 

Well 6

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

 

 2.5 nM abeta oligomer

Well 7

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

5 nM abeta oligomer 

Well 8

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

10 nM abeta oligomerStreptavidin

Well 9

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

Streptavidin

Beta-amyloid monomers25 nM abeta oligomer

Well 10

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP50

Streptavidin

Beta-amyloid oligomers nM abeta oligomer

Well 11

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

100 nM abeta oligomer 

Well 12

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

1000 nM abeta oligomer 

Well 13

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

0 nM abeta oligomer

Well 14

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

.1 nM abeta

Well 15

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

.25 nM abeta

Well 16

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

.5 nM abeta

Well 17

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

1 nM abeta

Well 18

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

2.5 nM abeta

Well 19

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

5 nM abeta

Well 20

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

10 nM abeta

Well 21

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

25 nM abeta

Well 22

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

50 nM abeta

Well 23

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

100 nM abeta

Well 24

HEK293

hEF1a: GmabL

hEF1a: GmabH

hEF1a: CD79A

hEF1a: CD79B

hEF1a: eYFP

1000 nM abeta

Expand
titleExpected Results

We expect to find clear membrane localization of red alexafluor conjugated streptaviding bound to beta amyloid bound to the BCR antibody. This will give us red halos around yellow cells with potentially blue nuclei due to DAPI staining. Moreover we expect a Kd on the order of 1 nM abeta based on the specifications of Gantenerumab. (According to the paper which introduced it, the Kd was 0.6 for plaques, 1.2 for oligomers, and 17 for monomers.) We are unsure whether the antibody will bind both monmerized and oligomerized beta amyloid.

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