Immunofluorescent analysis of BACE2 (green) showing staining in the cytoplasm of NIH-3T3 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a BACE2 polyclonal antibody (Product # PA1-754) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: C P(503) E V V N D E S S L V R H R W K(518)|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
PA1-754 detects human and mouse beta-secretase 2 (BACE2) from transfected cells. This product has recently been remanufactured in new rabbits as Cat. # PA1-1754.
PA1-754 has been successfully used in Western blot and ICC/IF procedures. By Western blot this antibody detects an ~58 kDa protein representing BACE2 from SY-SY5Y cells overexpressing the human gene.
The PA1-754 immunogen is amino acid residues 503-518 from human BACE2. This sequence is completely conserved in mouse. The PA1-754 immunizing peptide (Cat. # PEP-167) is available for use in neutralization and control experiments.
Amyloid beta peptide is the major constituent of amyloid plaques in the brains of individuals afflicted with Alzheimer and quote;s disease. This peptide is generated from the beta-amyloid precursor protein (beta APP) in a two-step process. The first step involves cleavage of the extracellular, amino-terminal domain of beta APP. Protein cleavage is performed by an aspartyl protease termed beta-secretase (BACE). This enzyme is synthesized as a propeptide that must be modified to the mature and active form by the prohormone convertase, furin. Beta APP cleavage by the mature form of BACE results in the cellular secretion of a segment of beta APP and a membrane-bound remnant. This remnant is then processed by another protease termed gamma-secretase. Gamma-secretase cleaves an intra-membrane site in the carboxyl-terminal domain of beta APP, thus generating the amyloid beta peptide. Gamma-secretase is believed to be a multi-subunit complex containing presenilin-1 and 2 as central components. Found associated with the presenilins is the transmembrane glycoprotein nicastrin. Nicastrin has been found to bind to the carboxyl-terminus of betaAPP and helps to modulate the production of the amyloid beta peptide.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Beta-secretase cleavage of Alzheimer's amyloid precursor protein by the transmembrane aspartic protease BACE.
PA1-754 was used in western blot to investigate beta-secretase cleavage of APP protein mediated by BACE
|Vassar R,Bennett BD,Babu-Khan S,Kahn S,Mendiaz EA,Denis P,Teplow DB,Ross S,Amarante P,Loeloff R,Luo Y,Fisher S,Fuller J,Edenson S,Lile J,Jarosinski MA,Biere AL,Curran E,Burgess T,Louis JC,Collins F,Treanor J,Rogers G,Citron M||Science (New York, N.Y.) (286:735)||1999|