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Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Acetylation

HDAC Activity Assay试剂盒(colorimetric) (ab1432)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (4)References (55)

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HDAC Activity Colorimetric Assay Kit Standard Curve
  • HDAC Activity Colorimetric Assay Kit Activity Assay

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric
  • Platform: Microplate (12 x 8 well strips)

概述

  • 产品名称

    HDAC Activity Assay试剂盒(colorimetric)
    参阅全部 HDAC 试剂盒
  • 检测方法

    Colorimetric
  • 检测类型

    Quantitative
  • 实验步骤

    Multiple steps standard assay
  • 种属反应性

    与反应: Mammals
  • 产品概述

    Inhibition of histone deacetylases (HDACs) has been implicated in the modulation of transcription and the induction of apoptosis or differentiation in cancer cells. However, screening HDAC inhibitory compounds has proven to be difficult over the past due to the lack of convenient tools for analyzing HDAC activity. The new Colorimetric HDAC Activity Assay Kit ab1432 provides a fast and convenient colorimetric method that eliminates radioactivity, extractions, or chromatography, as used in the traditional assays. The new method requires only two easy steps, both performed on the same microtiter plate. First, the HDAC colorimetric substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (e.g., HeLa nuclear extract or your own samples). Deacetylation of the substrate sensitizes the substrate, so that, in the second step, treatment with the Lysine Developer produces a chromophore. The chromophore can be easily analyzed using an ELISA plate reader or spectrophotometer. The assay is well suited for high throughput screening applications.

  • 说明

    This product is manufactured by BioVision, an Abcam company and was previously called K331 HDAC Activity Colorimetric Assay Kit. K331-100 is the same size as the 96 test size of ab1432.

    Read the entire protocol (attached to the datasheet) before beginning the procedure.

    The HeLa extract should be refrozen immediately at -20 or -70°C after each use to avoid loss of activity. The Lysine Developer should be refrozen immediately at –20 or -70°C after each use or aliquotted for future use.

    This kit contains enough reagents for about 100 tests. If positive and negative controls are included, the kit provides sufficient reagents for 5 positive control assays with the HeLa Nuclear Extract and 5 Negative Control assays with the HDAC Inhibitor, Trichostatin A.

  • 经测试应用

    适用于: Functional Studiesmore details
  • 平台

    Microplate (12 x 8 well strips)

性能

  • 存放说明

    Store at -80°C. Please refer to protocols.
  • 组件 标识符 96 tests
    10X HDAC Assay Buffer Green Cap 1 x 1ml
    Deacetylated Standard (10 mM) Yellow Cap 1 x 20µl
    HDAC Inhibitor (Trichostatin A, 1 mM) Blue Cap 1 x 10µl
    HDAC Substrate Amber Cap 1 x 500µl
    HeLa Nuclear Extract (5 mg/mL) Red Cap 1 x 50µl
    Lysine Developer Orange Cap 1 x 1ml
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Acetylation
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Acetylation
    • HDACs
    • Kits
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Cell Damage
  • 功能

    Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Deacetylates SP proteins, SP1 and SP3, and regulates their function. Component of the BRG1-RB1-HDAC1 complex, which negatively regulates the CREST-mediated transcription in resting neurons. Upon calcium stimulation, HDAC1 is released from the complex and CREBBP is recruited, which facilitates transcriptional activation. Deacetylates TSHZ3 and regulates its transcriptional repressor activity. Deacetylates 'Lys-310' in RELA and thereby inhibits the transcriptional activity of NF-kappa-B. Component a RCOR/GFI/KDM1A/HDAC complex that suppresses, via histone deacetylase (HDAC) recruitment, a number of genes implicated in multilineage blood cell development.
  • 组织特异性

    Ubiquitous, with higher levels in heart, pancreas and testis, and lower levels in kidney and brain.
  • 序列相似性

    Belongs to the histone deacetylase family. HD type 1 subfamily.
  • 翻译后修饰

    Sumoylated on Lys-444 and Lys-476; which promotes enzymatic activity. Desumoylated by SENP1.
    Phosphorylation on Ser-421 and Ser-423 promotes enzymatic activity and interactions with NuRD and SIN3 complexes. Phosphorylated by CDK5.
    Ubiquitinated by CHFR, leading to its degradation by the proteasome (By similarity). Ubiquitinated by KCTD11, leading to proteasomal degradation.
  • 细胞定位

    Nucleus.
  • Target information above from: UniProt accession Q13547 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 别名

    • HD1
    • HDAC1
    • HDAC1_HUMAN
    • Histone deacetylase 1

相关产品

  • Related Products

    • HDAC Activity Assay Kit (Fluorometric) (ab1438)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab1432于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Functional Studies
Use at an assay dependent concentration.
说明
Functional Studies
Use at an assay dependent concentration.

图片

  • HDAC Activity Colorimetric Assay Kit Standard Curve
    HDAC Activity Colorimetric Assay Kit Standard Curve
  • HDAC Activity Colorimetric Assay Kit Activity Assay
    HDAC Activity Colorimetric Assay Kit Activity Assay

    Different amount of nuclear extract (NE) were tested following kit protocol in the presence and absence of HDAC Inhibitor (Incubated for 4 hrs).

实验方案

  • HDAC Activity Assay Protocol (colorimetric)
  • Protocol Booklet

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (55)

发表研究结果有使用 ab1432?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab1432 被引用在 55 文献中.

  • Mu C  et al. Probiotics counteract hepatic steatosis caused by ketogenic diet and upregulate AMPK signaling in a model of infantile epilepsy. EBioMedicine 76:103838 (2022). PubMed: 35148983
  • Wei Y  et al. MicroRNA-874-3p promotes testosterone-induced granulosa cell apoptosis by suppressing HDAC1-mediated p53 deacetylation. Exp Ther Med 21:359 (2021). PubMed: 33732332
  • Du Y  et al. Suppression of HDAC2 by sodium butyrate alleviates apoptosis of kidney cells in db/db mice and HG-induced NRK-52E cells. Int J Mol Med 45:210-222 (2020). PubMed: 31746362
  • Nam MH  et al. Aspirin inhibits TGFß2-induced epithelial to mesenchymal transition of lens epithelial cells: selective acetylation of K56 and K122 in histone H3. Biochem J 477:75-97 (2020). PubMed: 31815277
  • Amnekar RV  et al. Histone deacetylase inhibitor pre-treatment enhances the efficacy of DNA-interacting chemotherapeutic drugs in gastric cancer. World J Gastroenterol 26:598-613 (2020). PubMed: 32103870
View all Publications for this product

客户评价及客户问答

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1-4 of 4 Abreviews or Q&A

Question

Can this product be used for mouse cell lysates? Also, how many samples can be read with this kit? Thank you!

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Abcam community

Verified customer

Asked on Jul 09 2010

Answer

Thank you for your enquiry. I can confirm that the kit (ab1432: HDAC Activity Assay K) will work with all mammalian samples and hence will work with mouse samples. You can perform 100 reactions with this kit. I hope this will be useful for you.

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Abcam Scientific Support

回复于 Jul 09 2010

Question

do you have a promotion price for a start-up lab? Thank you for your information.

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Abcam community

Verified customer

Asked on Mar 24 2005

Answer

Thank you for your email. I'm afraid that we are unable to provide discounts. If you have any additional questions, please let us know.

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Abcam Scientific Support

回复于 Mar 24 2005

Question

Which of these two HDAC activity assay kits is more sensitive? Do they detect HDAC3? How does the sensitivity compare to radioactive detection?

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Abcam community

Verified customer

Asked on Mar 24 2005

Answer

Thank you for your phone call. The originator of these two kits was able to provide me with the following information. Ab1438 (the fluorometric assay) is ~5 times more sensitive than ab1432 (the colorimetric assay). The HDAC activity assay detects all HDACs (including HDAC-3) that can deacetylate histone. A direct comparison with radioactive detection has not been done. Both ab1432 and ab1438 are currently out of stock and would take approximately 1-2 weeks to receive upon ordering. If you have any additional questions, please contact us again.

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Abcam Scientific Support

回复于 Mar 24 2005

Question

I have one question about the HDAC activity assay kit(ab1432). What is the lysis buffer recepi for preparing the lysate to measure the HDAC activity.Do I need to put proteinase inhibitor in it?

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Abcam community

Verified customer

Asked on Oct 15 2003

Answer

We would recommend using BioVision's Nuclear/Cytosol Fractionation kit (K266-100) to isolate Nuclear extract, and using such nuclear extract for the assays. Cell lysate can be prepared using Triton X-100 (1%) in Tris buffer (20 mM)+ 150 mM NaCl. However, the activity measured with cell lysate can be much lower in comparison the activity measured with nuclear extract.

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Abcam Scientific Support

回复于 Oct 20 2003

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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