Recombinant Human BACE-1 Protein, CF

Catalog #: 931-AS
9 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
931-AS-050

R&D Systems Recombinant Proteins and Enzymes

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Citations (9)

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human BACE-1 Protein, CF Summary

Product Specifications

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its ability to cleave a fluorogenic peptide substrate, Mca-SEVNLDAEFRK(Dpn)RR-NH₂ (Catalog # ES004). The specific activity is >3.5 pmol/min/µg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived human BACE-1 protein
Thr22-Tyr460 (pro) & Glu46-Tyr460 (mature), both with a C-terminal 10-His tag

Accession #

P56817.1

N-terminal Sequence
Analysis

Thr22 & Glu46

Structure / Form

Pro and Mature forms

Predicted Molecular Mass

50 kDa (Pro form) & 47 kDa (Mature form)

SDS-PAGE

68-70 kDa and 65-67 kDa, reducing conditions

Product Datasheets

931-AS

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

931-AS

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 500 μg/mL in sterile, deionized water.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 6 months from date of receipt, -20 to -70 °C as supplied. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Assay Procedure

Materials

Assay Buffer: 0.1 M Sodium Acetate, pH 4.0
Recombinant Human BACE -1 (rhBACE-1) (Catalog # 931-AS)
Fluorogenic Peptide Substrate IV: MCA-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Arg-Lys(DPN)-Arg-Arg-NH₂ (Catalog # ES004)
F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

Dilute rhBACE-1 to 20 ng/µL in Assay Buffer.
Dilute Substrate to 20 µM in Assay Buffer.
Load 50 µL of rhBACE-1 into a plate and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (RFU/min) x Conversion Factor** (pmol/RFU)
	amount of enzyme (µg)

*Adjusted for Substrate Blank
**Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:

rhBACE-1: 1 µg
Substrate: 10 µM

Reconstitution Calculator

Background: BACE-1

BACE-1 is an aspartic protease and an integral membrane protein (1-5). BACE-1 is the peptidase predominantly responsible for cleavage of the amyloid precursor protein beta site in the brain to generate the amyloid beta peptide. Because the amyloid beta peptide is a major component of amyloid plaques, BACE-1 has been implicated in the onset and/or progression of Alzheimer's disease. BACE-1 is expressed in a variety of human tissues. It is likely that this peptidase has functions in addition to its hydrolysis of the amyloid precursor protein. The peptidase activity of BACE-1 is optimal under mildly acidic conditions (pH 3.5-5.5), consistent with its proposed function in an acidic intracellular compartment.

References

Ermolieff, J. et al. (2000) Biochemistry 39:12450.
Lin, X. et al. (2000) Proc. Natl. Acad. Sci. USA 97:1456.
Sinha, S. et al. (1999) Nature 402:537.
Vassar, R. et al. (1999) Science 286:735.
Yan, R. et al. (1999) Nature 402:533.

Long Name

beta-site Ayloid Precursor Protein Cleaving Enzyme 1

Entrez Gene IDs

23621 (Human); 23821 (Mouse)

Alternate Names

APP beta-secretase; ASP2; Aspartyl protease 2; BACE1; BACE-1; BACEAsp 2; beta-secretase 1 precursor variant 1; beta-secretase 1; beta-site amyloid beta A4 precursor protein-cleaving enzyme; Beta-site amyloid precursor protein cleaving enzyme 1; Beta-site APP cleaving enzyme 1; beta-site APP-cleaving enzyme 1; beta-site APP-cleaving enzyme; EC 3.4.23; EC 3.4.23.46; FLJ90568; HSPC104; KIAA1149; memapsin-2; Membrane-associated aspartic protease 2; transmembrane aspartic proteinase Asp2

Related Research Areas

Citations for Recombinant Human BACE-1 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

9 Citations: Showing 1 - 9
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Identification and Analysis of Axolotl Homologs for Proteins Implicated in Human Neurodegenerative Proteinopathies
Authors: James, LM;Strickland, Z;Lopez, N;Whited, JL;Maden, M;Lewis, J;
Genes
Species: axolotl, Mouse
Sample Types: Protein
Applications: Bioassay
Dual Inhibitors of AChE and BACE-1 for Reducing Abeta in Alzheimer's Disease: From In Silico to In Vivo
Authors: N Stern, A Gacs, E Tátrai, B Flachner, I Hajdú, K Dobi, I Bágyi, G Dormán, Z L?rincz, S Cseh, A Kígyós, J Tóvári, A Goldblum
International Journal of Molecular Sciences, 2022-10-28;23(21):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay
Spatiotemporal processing of neural cell adhesion molecules 1 and 2 by BACE1 in vivo
Authors: W Kim, H Watanabe, S Lomoio, G Tesco
The Journal of Biological Chemistry, 2021-02-03;0(0):100372.
Species: Mouse
Sample Types: Tissue Homogenates
Applications: Bioassay
The botanical origin and antioxidant, anti-BACE1 and antiproliferative properties of bee pollen from different regions of South Korea
Authors: Y Zou, J Hu, W Huang, L Zhu, M Shao, C Dordoe, YJ Ahn, D Wang, Y Zhao, Y Xiong, X Wang
BMC Complement Med Ther, 2020-07-25;20(1):236.
Species: Human
Sample Types:
Applications: Enzyme Assay
Tissue-Specific Regulation of the Wnt/&beta-Catenin Pathway by PAGE4 Inhibition of Tankyrase
Authors: S Koirala, J Klein, Y Zheng, NO Glenn, T Eisemann, K Fon Tacer, DJ Miller, O Kulak, M Lu, DB Finkelstei, G Neale, H Tillman, P Vogel, DW Strand, L Lum, CA Brautigam, JM Pascal, WK Clements, PR Potts
Cell Rep, 2020-07-21;32(3):107922.
Species: Human
Sample Types: Whole Cells
Applications: Enzyme Assay
Gleevec shifts APP processing from a ?-cleavage to a nonamyloidogenic cleavage
Authors: WJ Netzer, K Bettayeb, SC Sinha, M Flajolet, P Greengard, V Bustos
Proc. Natl. Acad. Sci. U.S.A, 2017-01-23;0(0):.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay
Anti-amyloid precursor protein immunoglobulins inhibit amyloid-beta production by steric hindrance.
Authors: Thomas RS, Liddell JE, Kidd EJ
FEBS J., 2010-12-01;278(1):167-78.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay
BACE1 and BACE2 enzymatic activities in Alzheimer's disease.
Authors: Ahmed RR, Holler CJ, Webb RL
J. Neurochem., 2009-12-04;112(4):1045-53.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay
Inhibition of amyloid beta-peptide production by blockage of beta-secretase cleavage site of amyloid precursor protein.
Authors: Na CH, Jeon SH, Zhang G, Olson GL, Chae CB
J. Neurochem., 2007-06-01;101(6):1583-95.
Species: Human
Sample Types: Whole Cells
Applications: Enzyme Assay