Log In Sign up
The largest database of trusted experimental protocols

Hdac activity fluorometric assay kit

Manufactured by Abcam
Visit Supplier
Sourced in United States

The HDAC Activity Fluorometric Assay Kit is a tool used to measure the enzymatic activity of histone deacetylases (HDACs) in biological samples. It provides a fluorescence-based method for quantifying HDAC activity.

Automatically generated - may contain errors

Lab products found in correlation

Ms 275, by Cayman Chemical (3 mentions) Hif 1α transcription factor assay kit, by Cayman Chemical (2 mentions) L lactate assay kit, by Abcam (2 mentions) Sirtinol, by Cayman Chemical (2 mentions) Trichostatin a, by Merck Group (2 mentions) Vorinostat, by Cayman Chemical (2 mentions) Tubacin, by Cayman Chemical (2 mentions) Black 96 well plate, by Thermo Fisher Scientific (2 mentions) Synergy h1 hybrid reader, by Agilent Technologies (2 mentions) Hdac3 activity assay kit, by Abcam (1 mentions) Parp1 enzyme activity assay, by Merck Group (1 mentions) Sonic dismembrator model 100, by Thermo Fisher Scientific (1 mentions) Phosphatase inhibitor cocktail a, by Santa Cruz Biotechnology (1 mentions) Bradford assay, by Bio-Rad (1 mentions) Nuclear cytosol fractionation kit, by Abcam (1 mentions) Victor x3, by PerkinElmer (1 mentions) K266 25, by Abcam (1 mentions) Sigmaplot, by Grafiti LLC (1 mentions)

Spelling variants of this product

Fluorometric assay kit (63 citations) HDAC Activity Assay kit (11 citations) HDAC assay kit (4 citations) Fluorometric assays (3 citations) InSitu HDAC Activity Fluorometric Assay Kit (3 citations) Activity assay kits (2 citations) Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) (2 citations)

20 protocols using hdac activity fluorometric assay kit

1

Fluorometric HDAC Activity Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Two different types of in vitro HDAC activity assays were used (with or without cells). Test compounds were incubated with individual recombinant HDAC isoforms (HDAC 1-9) and the assay was performed according to the manufacturer's protocol (BPS Biosciences, USA). HT-29 cells were treated with MPT0G030 and SAHA for 24 h, and then total cell lysates were analyzed with Fluorometric HDAC Activity Assay Kit (BioVision, USA).
Wang L.T., Liou J.P., Li Y.H., Liu Y.M., Pan S.L, & Teng C.M. (2014). A novel class I HDAC inhibitor, MPT0G030, induces cell apoptosis and differentiation in human colorectal cancer cells via HDAC1/PKCδ and E-cadherin. Oncotarget, 5(14), 5651-5662.
+ Open protocol
+ Expand
2

Quantifying HDAC Activity in Kidney

Check if the same lab product or an alternative is used in the 5 most similar protocols
To determine the activity of HDAC, equal quantities of nuclear fraction proteins (20 μg) were extracted from kidney tissue (Active Motif, 40010), and analyzed using a Fluorometric HDAC Activity Assay Kit (BioVision, k330-100) as described previously (Wu et al., 2020 ).
Liu Y., Li Y.J., Loh Y.W., Singer J., Zhu W., Macia L., Mackay C.R., Wang W., Chadban S.J, & Wu H. (2021). Fiber Derived Microbial Metabolites Prevent Acute Kidney Injury Through G-Protein Coupled Receptors and HDAC Inhibition. Frontiers in Cell and Developmental Biology, 9, 648639.
+ Open protocol
+ Expand
3

HDAC Activity Assay in PBMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Global HDAC activity in PBMC nuclear extract was assessed using a Fluorometric HDAC Activity Assay Kit (Biovision, CA). All PBMC samples were incubated with HDAC substrate (Boc-Lys(Ac)-AMC). Deacetylation of substrate sensitizes substrate, and further addition of lysine developer produces a fluorophore, which was captured as the fluorescence read-out using a plate reader with Ex. = 350–380 nm and Em. = 440–460 nm settings. The standard curve was prepared as per recommended dilution range by the kit protocol. The absolute HDAC activity was calculated based on the slope determined by the standard deacetylated curve and expressed as μM/mL. Positive and negative controls were carried out during every experiment. Specific HDAC3 activity was assessed using the HDAC3 activity assay kit (Biovision, CA) as per the manufactures’ protocol. Absolute HDAC3 activity was calculated based on the slope determined by the standard deacetylated curve and expressed as pmol/min/mL.
Sathishkumar C., Prabu P., Balakumar M., Lenin R., Prabhu D., Anjana R.M., Mohan V, & Balasubramanyam M. (2016). Augmentation of histone deacetylase 3 (HDAC3) epigenetic signature at the interface of proinflammation and insulin resistance in patients with type 2 diabetes. Clinical Epigenetics, 8, 125.
+ Open protocol
+ Expand
4

Enzymatic Assays for Metabolic Markers

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Lactate in serum or the culture medium was measured with colorimetric L-Lactate Assay Kit (Abcam, Cambridge, MA, USA, #ab65331) according to the manufacturer’s instructions. HDAC activity was measured with the fluorometric HDAC Activity Assay kit (Abcam, #ab156064) according to the manufacturer’s instructions. HIF1α DNA binding activity in nuclear extracts was measured with HIF1α Transcription Factor Assay Kit (Cayman, Ann Arbor, MI, USA, #10006910) according to the manufacturer’s instructions. PKM2 activity was measured by a lactate dehydrogenase-coupled enzyme assay27 (link),60 (link). The assay was carried out with 1 µg of cell lysates with an enzyme buffer (50 mM tris-HCl [pH 7.5], 100 mM KCl, 10 mM MgCl2, 0.9 mM adenosine diphosphate, 0.6 mM phosphoenolpyruvate, 0.12 mM β-nicotinamide adenine dinucleotide and 4.8 U ml−1 lactate dehydrogenase). Enzyme activity can be measured at 340 nm absorbance by spectrophotometry.
Yang L., Xie M., Yang M., Yu Y., Zhu S., Hou W., Kang R., Lotze M., Billiar T.R., Wang H., Cao L, & Tang D. (2014). PKM2 Regulates the Warburg Effect and Promotes HMGB1 Release in Sepsis. Nature communications, 5, 4436.
+ Open protocol
+ Expand
5

Enzymatic Assays for Metabolic Markers

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Lactate in serum or the culture medium was measured with colorimetric L-Lactate Assay Kit (Abcam, Cambridge, MA, USA, #ab65331) according to the manufacturer’s instructions. HDAC activity was measured with the fluorometric HDAC Activity Assay kit (Abcam, #ab156064) according to the manufacturer’s instructions. HIF1α DNA binding activity in nuclear extracts was measured with HIF1α Transcription Factor Assay Kit (Cayman, Ann Arbor, MI, USA, #10006910) according to the manufacturer’s instructions. PKM2 activity was measured by a lactate dehydrogenase-coupled enzyme assay27 (link),60 (link). The assay was carried out with 1 µg of cell lysates with an enzyme buffer (50 mM tris-HCl [pH 7.5], 100 mM KCl, 10 mM MgCl2, 0.9 mM adenosine diphosphate, 0.6 mM phosphoenolpyruvate, 0.12 mM β-nicotinamide adenine dinucleotide and 4.8 U ml−1 lactate dehydrogenase). Enzyme activity can be measured at 340 nm absorbance by spectrophotometry.
Yang L., Xie M., Yang M., Yu Y., Zhu S., Hou W., Kang R., Lotze M., Billiar T.R., Wang H., Cao L, & Tang D. (2014). PKM2 Regulates the Warburg Effect and Promotes HMGB1 Release in Sepsis. Nature communications, 5, 4436.
+ Open protocol
+ Expand
6

HDAC Activity Assay of HCT116 Nuclear Extract

Check if the same lab product or an alternative is used in the 5 most similar protocols
HDAC activity of HCT116 nuclear extract was assayed using the Fluorometric HDAC Activity Assay Kit from Abcam (ab156064) according to the manufacturer’s protocol. Briefly, nuclear extract was prepared by dounce homogenizing cell pellets in lysis buffer (10 mM Tris-HCl, 10 mM NaCl, 3 mM MgCl2, and pH 7.4), then spinning out nuclei and resuspending in nuclear extraction buffer (50 mM HEPES, 420 mM NaCl, 0.5 mM EDTA, 0.1 mM EGTA, 10% glycerol, and pH 7.5). Protein concentration was determined by Bradford, and then 50 µg was added to dose curves of butyrate, propionate, and their corresponding acyl-CoAs. TSA was used as a positive control. Fluorescence signal (Ex/Em=360/450 nm) was read on a plate reader at 2 min intervals for 1 hr. Values were averaged over two technical replicates. IC50 values were calculated using GraphPad Prism.
Thomas S.P, & Denu J.M. (2021). Short-chain fatty acids activate acetyltransferase p300. eLife, 10, e72171.
+ Open protocol
+ Expand
7

Quantifying HDAC Activity in MM Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
The HDAC activity of MM cells was analyzed using the histone deacetylase (HDAC) Activity Assay Kit (Fluorometric) (Abcam, Cambridge, UK) according to the manufacturer’s instructions. Fluorescence intensity signals were measured using a fluorescence microplate reader (Ex/Em 355/460 nm).
Okabe S., Tanaka Y, & Gotoh A. (2021). Targeting phosphoinositide 3-kinases and histone deacetylases in multiple myeloma. Experimental Hematology & Oncology, 10, 19.
+ Open protocol
+ Expand
8

Evaluating HDAC and PARP1 Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
To assess HDACs and PARP1 activity, the day after plating, cells were exposed to MS-275 and harvested at different times. Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) (ab156064) from abcam was used to test HDACs activity and PARP1 Enzyme Activity Assay (17-10149) was from Sigma-Aldrich (St. Louis, MO, USA) to test PARP1 activity. Assays were performed accordingly with the manufacturer’s instructions.
Cassandri M., Pomella S., Rossetti A., Petragnano F., Milazzo L., Vulcano F., Camero S., Codenotti S., Cicchetti F., Maggio R., Festuccia C., Gravina G.L., Fanzani A., Megiorni F., Catanoso M., Marchese C., Tombolini V., Locatelli F., Rota R, & Marampon F. (2021). MS-275 (Entinostat) Promotes Radio-Sensitivity in PAX3-FOXO1 Rhabdomyosarcoma Cells. International Journal of Molecular Sciences, 22(19), 10671.
+ Open protocol
+ Expand
9

HDAC Inhibition Assay in HCT116 Cells

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
HCT116 cells were seeded in 10-cm dish and treated with indicated concentrations of compound 11 and SAHA for 24 h, and then cell lysates were subjected to a HDAC Fluorometric Activity Assay Kit (K330–100, Biovision Inc.) as described previously [32 (link)].
Chen C.H., Lee C.H., Liou J.P., Teng C.M, & Pan S.L. (2015). Molecular mechanisms underlying the antitumor activity of (E)-N-hydroxy-3-(1-(4-methoxyphenylsulfonyl)-1,2,3,4-tetrahydroquinolin-6-yl)acrylamide in human colorectal cancer cells in vitro and in vivo. Oncotarget, 6(34), 35991-36002.
+ Open protocol
+ Expand
10

HDAC Activity Assay for Anticancer Drugs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were plated in 10-cm dish (1×106 cells/ml) and treated with different doses of MPT0E028 and SAHA for 24 h, and then cell lysate was subjected to a HDAC Fluorometric Activity Assay Kit (K330-100, Biovision Inc.) to determine the HDAC activity. 50 μg of cell lysate was added to react with fluorometric substrate t-butoxycarbonyl-Lys (AC)-amido-4-methylcoumarin (Boc-Lys (AC) AMC) and incubated at 37°C for 30 minutes. After the incubation, the lysine developer in this kit was added and the mixture was incubated for another 30 min at 37°C. Fluorescence was detected by plate reader paradigm detection platform (Beckman coulter) with Ex=360 (excitation) and Em=465 nm (emission).
Huang H.L., Peng C.Y., Lai M.J., Chen C.H., Lee H.Y., Wang J.C., Liou J.P., Pan S.L, & Teng C.M. (2014). Novel oral histone deacetylase inhibitor, MPT0E028, displays potent growth-inhibitory activity against human B-cell lymphoma in vitro and in vivo. Oncotarget, 6(7), 4976-4991.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!