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BACE-1 Overexpression Lysate

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Western Blot: BACE1 Overexpression Lysate (Adult Normal) [NBP2-04325] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for BACE1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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BACE-1 Overexpression Lysate Summary

Description

BACE-1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC209115

Accession#: NM_012104

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Gene
BACE1

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Theoretical MW
51 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for BACE-1 Overexpression Lysate

  • 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

Background

BACE1 is a protease responsible for the proteolytic processing of the amyloid precursor protein (APP). Amyloid beta peptide is the major constituent of amyloid plaques in the brains of individuals afflicted with Alzheimers 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), of which there are two isoforms, BACE1 and 2. 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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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Bioinformatics

Gene Symbol BACE1