Page History

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Upon detection of beta-amyloid plaques (by receptor protein binding or by miRNA sensing) a transcription factor is cleaved from the receptor protein (LilrB2 or PirB) to then promote expression of a miRNA as well as BACE2. This miRNA, in turn, down-regulates BACE1.

Project PlanTimeline

TO DO

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Experiments

Lab Notebook

Experiment	Expected Results	Controls	Protocols	Parts	Completed?
Demonstrate inducible upregulation of BACE2 with GFP see more image



Perform a functional assay of inducible upregulation of BACE2

Purchases

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Experiments

Seeding 24-well Plate......Transfection......FACS

Experiment

Demonstrate effectiveness of miRNA in down-regulating BACE1

transfection plans

Demonstrate inducible upregulation of BACE2-YFP

FRET

Purchases

Item	Link
Item	Link	Purchased?
BACE2	http://dnasu.org/DNASU/GetCloneDetail.do?cloneid=76140		BACE1 (mouse)	https://www.addgene.org/26736/
BACE1 (human)	http://dnasu.org/DNASU/GetCloneDetail.do?cloneid=78236

Options for Directly Assaying BACE2 Expression

This is a kit to directly measure BACE2 expressionsigmaaldrichcatalog/search?interface=All&term=bace2&N=0+14574647&mode=match+partialmax&focus=product&lang=en&region=US

Link	Cost	Comments
Bace2 activity assay
https://www.anaspec.com/products/product.asp?id=57836	$435
Bace1 activity assay	http://www.	anaspec.com/	$315-$380	This is a kit to directly measure BACE2 expression
http://www.sciencedirect.com.ezproxyberklee.flo.org/science/article/pii/S1550413111003019; http://www.novusbio.com/gene-id/57393#fq=category%3A%22Peptides%20and%20Proteins%22	$369 for 10ug of TMEM27	This assay would involve measuring Tmem27 cleavage by Bace2
http://www.pnas.org/content/97/17/9712.long	n/a	This assay would involve measuring A-beta cleavage by Bace2
http://www.researchgate.net/post/Which_are_the_dye_suitable_for_visualization_of_neuron_with_beta-amyloid_protein_and_plaque10
atomic force microscopy, Pittsburgh compound B