Cleavage of amyloid precursor protein (APP) into 40 or 42 amino acid amyloid beta (ab) peptides is a result of the action of two proteases beta- and gamma-secretase. The beta-site APP cleaving enzyme, or BACE1, is a transmembrane aspartic protease that is localized in the Golgi and endosomes. Low levels of BACE1 mRNA are found in most adult tissues with a higher level of expression in pancreas.
Applications:
Suitable for use in Immunohistochemistry, Flow Cytometry, Western Blot and Direct ELISA. Other applications not tested.
Recommended Dilutions:
Immunohistochemistry: 8ug/ml detects BACE1 in paraffin-embedded human Alzheimer�s brain tissue sections.
Flow Cytometry: 25ug/ml; 10ul labels 1-2.5x10e5 cells in a total reaction volume < 200ul. The binding of unlabeled monoclonal antibodies may be visualized by adding 10ul of a 25ug/ml stock solution of a secondary developing reagent such as goat anti-mouse IgG conjugated to a fluorochrome.
Western Blot: 2ug/ml detects human BACE1 in cell or tissue extracts under reducing conditions only.
Direct ELISA: 0.5-1.0ug/ml. The detection limit is ~1ng/well.
Optimal dilutions to be determined by the researcher.
Western Blot Buffers:
Blotting Buffer: 25mM Tris, pH 7.4, 0.15M sodium chloride, 0.05% Tween-20
Blocking Solution: 5% nonfat dry milk in Blotting Buffer
Antibody Solution: 5% nonfat dry milk in Blotting Buffer
Storage and Stability:
Lyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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