Beta Secretase Activity Assay试剂盒(Fluorometric) (ab65357)

概述

  • 产品名称
    Beta Secretase Activity Assay试剂盒(Fluorometric)
    参阅全部 Beta secretase 试剂盒
  • 样品类型
    Urine, Serum, Plasma, Other biological fluids, Cell Lysate, Cell culture media, Tissue Lysate
  • 检测类型
    Enzyme activity
  • 检测时间
    1h 00m
  • 种属反应性
    与反应: Other species, Mammals
  • 产品概述

    Beta Secretase Activity Assay Kit (ab65357) is a convenient fluorescence method for detecting beta secretase activity in biological and purified samples. The assay uses a secretase-specific peptide conjugated to two reporter molecules, EDANS and DABCYL. In the uncleaved form, the fluorescent signal from EDANS is quenched by the physical proximity of the DABCYL moiety. Cleavage of the peptide by beta secretase physically separates EDANS and DABCYL allowing for the release of a fluorescent signal. The level of secretase enzymatic activity in samples is proportional to the level of fluorescence intensity.
    Visit our FAQs page for tips and troubleshooting.

  • 说明

    β-Secretase (Beta secretase) is responsible for the proteolytic process of the amyloid precursor protein (APP). It cleaves APP at the N-terminus of the A-beta peptide sequence, between residues 671 – 672, leading to the generation and extracellular release of beta-cleaved soluble APP, and a corresponding cell-associated C-terminal fragment which is later released by gamma-secretase. It has been implicated to be an excellent target for anti-amyloid therapy for the treatment of Alzheimer’s disease

性能

  • 存放说明
    Store at -20°C. Please refer to protocols.
  • 组件 标识符 100 tests
    Active ß -Secretase Red 1 x 20µl
    ß -Secretase Extraction Buffer NM 1 x 25ml
    ß -Secretase Inhibitor (in DMSO) Blue 1 x 10µl
    ß -Secretase Reaction Buffer (2X) WM 1 x 10ml
    ß -Secretase Substrate (in DMSO) Amber NM 1 x 200µl
  • 研究领域
  • 相关性
    Beta secretase 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. Beta APP cleavage by beta-secretase 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.
  • 细胞定位
    Membrane; Single-pass type I membrane protein.
  • 别名
    • APP beta secretase
    • ASP 2
    • ASP2
    • BACE
    • BACE1
    • Beta secretase 1
    • Beta site APP cleaving enzyme 1
    • Memapsin2
    see all

图片

  • Beta Secretase Activity measured in biological fluids. Tested neat. 

  • Beta Secretase Activity measured in tissue lysates (each with protein concentration of 4 mg/mL)

  • Beta Secretase Activity measured in cell lysates (each with concentration of 2e7 cells/mL).

  • Beta-Secretase Cleavage Activity assayed using ab65357.

实验方案

文献

This product has been referenced in:
  • Do Carmo S  et al. Rescue of Early bace-1 and Global DNA Demethylation by S-Adenosylmethionine Reduces Amyloid Pathology and Improves Cognition in an Alzheimer's Model. Sci Rep 6:34051 (2016). Read more (PubMed: 27681803) »
  • Blondrath K  et al. The nuclear cofactor receptor interacting protein-140 (RIP140) regulates the expression of genes involved in Aß generation. Neurobiol Aging 47:180-191 (2016). Functional Studies . Read more (PubMed: 27614112) »

See all 10 Publications for this product

客户评价及客户问答

Abreviews
β-secretase activity in the mouse brain were measured using this kit. Solubilized membranes were extracted from brain tissues using β-secretase extraction buffer in this kit.
Username

Ms. Ji Hye Han

Verified customer

提交于 Jun 27 2018



We recommend to use 2 – 3x volume of ice-cold extraction buffer (provided with the kit) for tissue samples and to homogenize it on ice. The composition of the extraction buffer is proprietary information but I am happy to confirm that i...

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I contacted the laboratories. This kit has not been tested with DMSO dissolved samples. However, I am sorry it is most likely that samples in DMSO will not work well since enzymes are sensitive to DMSO. Typically >1-2% DMSO is not recommended...

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The buffer used might work but we would strongly suggest using the extraction buffer provided in the kit for the best results. The best way would be to store cell lysates at -80C if needed. The number of cells mentioned in the datasheet are b...

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Thank you for contacting us. This kit does allow you to measure BACE1 from cells without isolating the enzyme. We have tested this protocol and made the protocol from use with cells that were lysed using the kit's extraction buffer.

The public...

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Since this is an enzymatic assay, I would not recommend using a buffer containing EDTA.

Yes, you can add reaction buffer to the substrate. So if you are adding 18 ul of the buffer to 2 ul of the substrate, add only 32 ul of the buffer in the ...

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Tips to ensure maximum beta secretase activity include:
1. Ensure assay buffer at room temperature
2. Perhaps protein in your sample is inhibiting somehow beta secretase activity. To test this, try spiking active beta secretase cont...

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wenn nach der Beratung mit ihrem Chef weitere Fragen aufkämmen können sie sich gerne nocheinmal melden.

Ich wünsche ihnen weiterhin viel Erfolg mit ihren Experimenten

Vielen Dank für ihre Anfrage.

Für den Versuchsaufbau dieses Kits sollten in jedem Fall immer die positiv und die negativ Kontrollen sowie verschiedene Konzentrationen der Proben gemessen werden. Wenn Sie ein geeignetes Gerät ver...

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Thank you for contacting us with your questions about ab65357.

Do you know what the detection bandwidth is on your machine? If the bandwidth is ±20, you can use this wavelength, but if not, then unfortunatelythe difference inwavelength...

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1-10 of 17 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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