Histone Deacetylase (HDAC) Activity Assay试剂盒(Fluorometric) (ab156064)
Key features and details
- Assay type: Enzyme activity
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Cell culture extracts, Tissue Extracts
概述
-
产品名称
Histone Deacetylase (HDAC) Activity Assay试剂盒(Fluorometric)
参阅全部 HDAC1 试剂盒 -
检测方法
Fluorescent -
样品类型
Cell culture extracts, Tissue Extracts -
检测类型
Enzyme activity -
检测时间
1h 00m -
种属反应性
与反应: Mammals -
产品概述
Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) ab156064 detects HDAC activity in lysates.
The HDAC Activity Assay Kit is primarily designed for the evaluation of HDAC inhibitors using a crude HDAC fraction. Also, any cultured primary cell, cell line, or tissue homogenate can be assayed for HDAC activity with the kit if the appropriate dose of HDAC specific inhibitor e.g. Trichostatin A is used.
The HDAC activity assay protocol is based on an acetylated peptide which is conjugated to AMC. AMC is a fluorescent dye and its fluoresence is quenched when conjugated to the peptide. When the HDAC de-acetylates the peptide, it becomes susceptible to cleavage by an enzyme (the Developer component). This then releases free AMC, which can be measured using a fluorescence microplate reader (Ex/Em 355/460 nm).
The HDAC assay has been shown to detect the activity of the HDAC family, at least class I HDACs in Human or animal cell lysates or in column fractions. The assay shows good linearity of sample response. The assay may be used to follow the purification of HDACs or may be used to detect the presence of HDACs in cell lysates.
-
说明
Applications for this kit include:
1. Monitoring the purification of HDACs including HDAC1, 2, 3 and 8.
2. Screening inhibitors or activators of HDACs.
3. Detecting the effects of pharmacological agents on HDACs.Background information on HDACs
Histone Deacetylases (HDACs) are a class of enzymes responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4), allowing the histones to wrap the DNA more tightly.
HDAC proteins occur in four groups (class I, class IIA, class IIB, class III, class IV) based on function and DNA sequence similarity.
Classes I, IIA and IIB are considered "classical" HDACs whose activities are inhibited by trichostatin A (TSA), whereas class III is a family of NAD+-dependent proteins (sirtuins) not affected by TSA. Class IV is considered an atypical class on its own, based solely on DNA sequence similarity to the others. -
平台
Microplate reader
性能
-
存放说明
Please refer to protocols. -
组件 100 tests Crude HDAC 1 x 500µl Developer 1 x 500µl Fluoro-Deacetylated Peptide (0.2 mM) 1 x 100µl Fluoro-Substrate Peptide (0.2 mM) 1 x 500µl HDAC Assay Buffer 2 x 1ml HDAC Stop Solution 2 x 1ml Trichostatin A (10µM) 1 x 500µl -
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. -
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.
Ubiquitinated by CHFR, leading to its degradation by the proteasome. -
细胞定位
Nucleus. - Information by UniProt
-
别名
- DKFZp686H12203
- GON 10
- HD1
see all
图片
-
Butyrate, propionate, and TSA inhibited HDACs activity in HUVEC. HDACs activity were inhibited in HUVEC by treatment of butyrate and propionate. The results were normalized using the control as 100%.
-
8 uL (corresponding to 6.7e6 cells) of nuclear extracts (ab113474) from HL60 cells were assessed kinetically (+/- SD).
-
Different volumes of positive control, with inhibitor (TA), kinetically read (+/- SD).
-
Measurement of HeLa cell endogenous HDAC1 in an immunoprecipitated sample with anti-HDAC1 antibody and following the Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) (ab156064) protocol.
-
Measurement of HeLa cell endogenous HDAC2 in an immunoprecipitated sample with anti-HDAC2 antibody following the Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) (ab156064) protocol.
数据表及文件
-
SDS download
-
Datasheet download
文献 (36)
ab156064 被引用在 36 文献中.
- Jiménez-Alcántar P et al. PaDef (Persea americana var. drymifolia), a Plant Antimicrobial Peptide, Triggers Apoptosis, and Induces Global Epigenetic Modifications on Histone 3 in an Acute Lymphoid Leukemia Cell Line. Front Mol Biosci 9:801816 (2022). PubMed: 35141282
- Taiarol L et al. Givinostat-Liposomes: Anti-Tumor Effect on 2D and 3D Glioblastoma Models and Pharmacokinetics. Cancers (Basel) 14:N/A (2022). PubMed: 35740641
- Yuan S et al. The histone modification reader ZCWPW1 promotes double-strand break repair by regulating cross-talk of histone modifications and chromatin accessibility at meiotic hotspots. Genome Biol 23:187 (2022). PubMed: 36068616
- Hammer SS et al. Fasting and fasting-mimicking treatment activate SIRT1/LXRa and alleviate diabetes-induced systemic and microvascular dysfunction. Diabetologia 64:1674-1689 (2021). PubMed: 33770194
- Lee IS et al. Effects of 4-Hexylresorcinol on Craniofacial Growth in Rats. Int J Mol Sci 22:N/A (2021). PubMed: 34445640