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BACE1 cell lysate

Human BACE1 HEK293 Overexpression Lysate

Gene Names
BACE1; ASP2; BACE; HSPC104
Applications
Western Blot
Synonyms
BACE1; Human BACE1 HEK293 Overexpression Lysate; Human BACE1/ASP2 HEK293 Cell Lysate (WB positive control); beta-site APP-cleaving enzyme 1; Human ASP2 Overexpression Lysate; Human BACE Overexpression Lysate; Human HSPC104 Overexpression Lysate; BACE1 cell lysate
Ordering
For Research Use Only!
Host
HEK293 Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
446aa
Sequence
Met1-Thr457
Applicable Applications for BACE1 cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Human
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the extracellular domain (amino acid residue Met 1-Thr 457) of human BACE1 (NP_036236.1) precursor was expressed with a C-terminal polyhistidine tag.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for BACE1 cell lysate
Beta-site APP-cleaving enzyme 1 (BACE1) is an aspartic-acid protease important in the formation of myelin sheaths in peripheral nerve cells. In the brain, This protein is expressed highly in the substantia nigra, locus coruleus and medulla oblongata. Strong BACE1 expression has also been described in pancreatic tissue. BACE1 has a pivotal role in the pathogenesis of Alzheimer's disease. In Alzheimer's disease patients, BACE1 levels were elevated although mRNA levels were not changed. It has been found that BACE1 gene expression is controlled by a TATA-less promoter. The translational repression as a new mechanism controlling its expression. And the low concentrations of Ca(2+) (microM range) significantly increased the proteolytic activity of BACE1. Furthermore, BACE1 protein is ubiquitinated, and the degradation of BACE1 proteins and amyloid precursor protein processing are regulated by the ubiquitin-proteasome pathway. It has also been identified as the rate limiting enzyme for amyloid-beta-peptide (Abeta) production.

This Human BACE1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of BACE1 protein from the overexpression lysate was verified.
References
Christensen MA, et al. (2004) Transcriptional regulation of BACE1, the beta-amyloid precursor protein beta-secretase, by Sp1. Mol Cell Biol. 24(2):865-74.Stockley JH, et al. (2007) The proteins BACE1 and BACE2 and beta-secretase activity in normal and Alzheimer's disease brain. Biochem Soc Trans. 35(Pt 3): 574-6.Savonenko AV, et al. (2008) Alteration of BACE1-dependent NRG1/ErbB4 signaling and schizophrenia-like phenotypes in BACE1-null mice. Proc Natl Acad Sci U S A. 105(14): 5585-90.Hayley M, et al. (2009) Calcium enhances the proteolytic activity of BACE1: An in vitro biophysical and biochemical characterization of the BACE1-calcium interaction. Biochim Biophys Acta. 1788(9): 1933-8.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
42,107 Da
NCBI Official Full Name
beta-secretase 1 isoform A preproprotein
NCBI Official Synonym Full Names
beta-site APP-cleaving enzyme 1
NCBI Official Symbol
BACE1
NCBI Official Synonym Symbols
ASP2; BACE; HSPC104
NCBI Protein Information
beta-secretase 1; asp 2; memapsin-2; APP beta-secretase; aspartyl protease 2; beta-site APP cleaving enzyme 1; beta-secretase 1 precursor variant 1; transmembrane aspartic proteinase Asp2; membrane-associated aspartic protease 2; beta-site amyloid beta A4
UniProt Protein Name
Beta-secretase 1
Protein Family
UniProt Gene Name
BACE1
UniProt Synonym Gene Names
BACE; KIAA1149; ASP2; Asp 2
UniProt Entry Name
BACE1_HUMAN

Uniprot Description

BACE: an integral membrane glycoprotein and aspartic protease that is found mainly in the Golgi. Cleaves APP at the amino terminus of the A-beta peptide sequence, leading to the generation and extracellular release of beta-cleaved soluble APP, and a corresponding cell-associated carboxy-terminal fragment which is later release by gamma-secretase. Four splice-variant isoforms have been described.

Protein type: Protease; EC 3.4.23.46; Membrane protein, integral

Chromosomal Location of Human Ortholog: 11q23.2-q23.3

Cellular Component: multivesicular body; Golgi apparatus; cell surface; cytoplasmic vesicle membrane; integral to plasma membrane; axon; endoplasmic reticulum; late endosome; integral to membrane; plasma membrane; trans-Golgi network; endosome

Molecular Function: protein binding; enzyme binding; beta-amyloid binding; beta-aspartyl-peptidase activity; aspartic-type endopeptidase activity

Biological Process: beta-amyloid metabolic process; membrane protein ectodomain proteolysis; proteolysis

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Product Notes

The BACE1 bace1 (Catalog #AAA8113427) is a Cell Lysate produced from HEK293 Cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 446aa. AAA Biotech's BACE1 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the BACE1 bace1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Thr45 7. It is sometimes possible for the material contained within the vial of "BACE1, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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