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HDAC Activity Assay试剂盒(Fluorometric) (ab1438)

Submit a review Q&A (9)References (5)
HDAC Activity Assay Kit

    Key features and details

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

    概述

    • 产品名称

      HDAC Activity Assay试剂盒(Fluorometric)
      参阅全部 HDAC 试剂盒

    • 检测方法

      Fluorescent

    • 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 deacetylase (HDAC) has been implicated in the modulation of transcription and induction of apoptosis or differentiation in cancer cells.


      However, screening of compounds for HDAC inhibition has been difficult due to the lack of convenient tools for analyzing HDAC activity.


      The new Fluorometric HDAC Activity Assay Kit provides a fast and fluorescence-based method that eliminates radioactivity, extractions, or chromatography, as used in traditional assays.


      The new procedure requires only two easy steps, both performed on the same microtiter plate.


      First, the HDAC fluorometric substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (e.g., HeLa nuclear extract or others).


      Deacetylation of the substrate sensitizes the substrate, so that, in the second step, treatment with the Lysine Developer produces a fluorophore.


      The fluorophore can be easily analyzed using a fluorescence plate reader or a fluorometer.


      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 K330 HDAC Activity Fluorometric Assay Kit. K330-100 is the same size as the 96 test size of ab1438.

      Read the entire protocol before beginning the procedure.

      The HeLa extract should be refrozen immediately at –70°C after each use to avoid loss of activity.

      If positive and negative controls are designed, 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.

    • 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 (4 mM) Yellow cap 1 x 20µl
      HDAC Inhibitor (Trichostatin A, 1mM) Blue Cap 1 x 10µl
      HDAC Substrate Amber cap 1 x 500µl
      HeLa Nuclear Extract (5 mg/ml) Red cap 1 x 10µl
      Lysine Developer Orange cap 1 x 1ml

    • 研究领域

    • 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.
    • Information by UniProt

    • 别名

      • HD1
      • HDAC1
      • HDAC1_HUMAN
      • Histone deacetylase 1

    图片

    • HDAC Activity Assay Kit
      HDAC Activity Assay Kit

      Typical standard curve using ab1438.

    文献 (5)

    发表研究结果有使用 ab1438？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab1438 被引用在 5 文献中.

    • Perez M  et al. Tau--an inhibitor of deacetylase HDAC6 function. J Neurochem 109:1756-66 (2009). PubMed: 19457097
    • Li HL  et al. Curcumin prevents and reverses murine cardiac hypertrophy. J Clin Invest 118:879-93 (2008). PubMed: 18292803
    • Cheung WL  et al. Acetylation and chromosomal functions. Curr Opin Cell Biol 12:326-33 (2000). PubMed: 10801466
    • Strahl BD & Allis CD The language of covalent histone modifications. Nature 403:41-5 (2000). PubMed: 10638745
    • Grunstein M Histone acetylation in chromatin structure and transcription. Nature 389:349-52 (1997). PubMed: 9311776

    客户评价及客户问答

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    1-9 of 9 Abreviews or Q&A

    Question

    We have theHDAC Activity Assay Kit (Fluorometric) (ab1438): Abcambut the protocol for this is lost. There is no
    information regarding this on website.

    I kindly request you to please send me the protocol with expiry date of this kit as soon as possible.

    Kit details:
    LOT ****

    HDAC Activity Assay Kit (Fluorometric) (ab1438)

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    Answer

    Thank you for contacting us.

    Please find the assay protocol for this kit attached. The lot you specified expired in December 2011, and would no longer be covered under our Abpromise guarantee. It is possible that the reagents may still be working, but this would need to be tested. I hope this helps, please let me know if you need any additional information.

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    Question

    how many assays can be run with 1 kit?

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    Answer

    1.) The total number of test wells which can be performed with this kit is 100 (some of those test wells will then be occupied for using the positive and negative controls as well as the standards and blanks) 2.) There is enough positive and negative control to run 5 separate experiments (e.g. 5 times 20 wells in total then)

    Read More

    Question

    Does this kit have any species specificity? Can I use it for both human and rat samples?

    Read More
    Answer

    Thank you for your inquiry. This assay kit can be used with mammalian species, so yes it can be used with the indicated samples. I hope this information helps. Please contact us with any other questions.

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    Question

    Le kit ab1438 marchera t-il sur des lysats totaux, ou faut-il faire une extraction nucleaire? -Will the kit ab1438 work on total lysates or only nuclear lysates?

    Read More
    Answer

    Voici la reponse du laboratoire: "It depends on the user's purpose. There may be some enzymes in the cytosol that may interfere with the reaction results if the cell extracts were used. That is why we suggest the use of the nuclear extract." J'espere que cette reponse vous aidera.

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    Question

    We are trying to test our compounds on inhibition of HDAC enzyme using your HDAC fluorimetric assay kit. However, there is an interaction between our compounds and the HDAC assay buffer. We would appreciate your sending us a list of components of the HDAC assay buffer. Thank you. Yours,

    Read More
    Answer

    Thank you for your enquiry. The buffer contains Tris, NaCl, KCl, and MgCl2. I hope this information helps. Please do not hesitate to contact us again if you need anything further.

    Read More

    Question

    Thank you for your help. Can I ask two more questions? Do you know how to prepare the nuclear extract for HDAC assay?? When we measure fluorescence plate reader, we only can use Ex and Em? If our machine does not have this function, what I have to do? Thank you very much.

    Read More
    Answer

    Here is a protocol for getting a nuclear extract that will work for the HDAC activity assay: https://www.abcam.com/assets/pdf/protocols/Nuclear%20fractionation%20protocol.pdf I do not know what kind of plate reader you have, but if it cannot measure fluorescence, you should get a colorimetric kit for the HDAC assay, ab1432: https://www.abcam.com/HDAC-Activity-Assay-Kit-colorimetric-ab1432.html I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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    Question

    We ordered HDAC activity assay kit (ab 1438) from you. We set two wave length L1 (360 nm) and L2 (450 nm). Next is what we got: 1 2 3 4 5 6 7 8 9 0.000 0.132 0.017 0.041 0.02 0.036 0.018 0.040 0.027 0.000 0.006 0.038 0.046 0.044 0.056 0.043 0.049 0.012 #1 is blank, #2 is positive control from the kit. #3-#9 are our samples treated with different conditions and we would like to compare the difference between them. Could you please tell me how we should calculate the relative activity of our samples from the two values? Should we divide them or is there any formula to calculate it? We greatly appreciate your answer!

    Read More
    Answer

    Thank you for your email. You want to read and record the fluorescence as measured at an excitation (Ex.) of 350-380 nm and emission (Em.) of 440-460 nm. You can simply express the results as Relative fluorescence units/ug protein sample. The originator of this kit has said that "These values seem to be really low for fluorometric assay. Please check the machine setting to make sure it's correct." If you have any additional questions, please let us know.

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    Question

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

    Read More
    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.

    Read More

    Question

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

    Read More
    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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