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ENZO熱銷產(chǎn)品——FLUOR DE LYS? HDAC fluorometric activity assay kit組蛋白去乙?;笩晒饣钚詸z測試劑盒

2024-03-12
瀏覽次數(shù): 54

ENZO熱銷產(chǎn)品——FLUOR DE LYS??HDAC fluorometric activity assay kit組蛋白去乙酰化酶熒光活性檢測試劑盒


目前,組蛋白去乙?;敢种苿┮呀?jīng)顯示出其作為抗腫瘤藥物的前景,這也自然激發(fā)了研究者們對于篩選HDAC(Histone deacetylase,組蛋白去乙酰化酶)抑制化合物的興趣。Enzo的FLUOR DE LYS?HDAC熒光活性檢測試劑盒(貨號:BML-AK500-0001)是一種靈敏、便捷的組蛋白去乙?;笝z測試劑盒,可替代使用傳統(tǒng)的放射性標記的乙酰化組蛋白和肽/HPLC法檢測組蛋白去乙?;傅姆桨?。


FLUOR DE LYS?去乙?;笝z測平臺是Enzo的升級酶活性檢測系列產(chǎn)品的基礎,其中的產(chǎn)品就包括用于篩選HDAC(組蛋白去乙?;福┗钚哉{(diào)節(jié)劑和Sirtuin(長壽蛋白,一種去乙?;福┗钚哉{(diào)節(jié)劑的試劑盒。Enzo還可提供基于熒光共振能量轉移(FRET)的試劑盒,用以檢測基質(zhì)金屬蛋白酶(MMPs)和天冬氨酸特異性半胱氨酸蛋白酶(Caspase)的活性,以及提供基于磷酸特異性抗體的激酶檢測試劑盒。


這些檢測證明了Enzo在組裝小肽、合成活性重組蛋白、熒光標記和檢測技術,以及抗體開發(fā)等多方面協(xié)同的能力,也保障了Enzo能夠生產(chǎn)出功能強大且重復性好的檢測試劑盒。

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Enzo的酶活性檢測試劑盒均具有以下特點:

高品質(zhì):高活性、高純度的重組酶,可呈現(xiàn)高質(zhì)量的底物結合效果

靈活性:批量酶和底物可用于篩選后研究

強創(chuàng)新:多種readout?格式的專利底物/顯影劑化學物質(zhì)

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作用機制:

FLUOR DE LYS??HDAC fluorometric activity assay kit基于Enzo特有的?FLUOR DE LYS?(熒光法組蛋白去乙酰化酶-賴氨酸)底物和顯影劑組合,只需2步簡單的混合步驟,即可在同一個96?孔板上進行檢測。首先,將含有乙?;嚢彼醾孺湹腇LUOR DE LYS?底物與含有HDAC活性的樣品(HeLa核或其他提取物、純化酶、珠結合免疫復合物等)一起孵育。底物的去乙酰化會使底物增敏,所以在第二步中,將其與FLUOR DE LYS?顯影劑混合后會產(chǎn)生熒光團。


該檢測方法目前已經(jīng)成功用于所有已知的I類HDAC——HDAC1、HDAC2、HDAC3和HDAC8(見產(chǎn)品數(shù)據(jù)表)和II?類HDAC 4-7、9和10,以及人類Sir2同源物和SIRT1(見產(chǎn)品數(shù)據(jù)表)的制備。Enzo Life Sciences的研究表明,F(xiàn)LUOR DE LYS?底物具有細胞滲透性,并可在原位被細胞HDAC去乙?;C撘阴;牡孜镌诩毎麅?nèi)積聚后,可通過向細胞裂解液中加入FLUOR DE LYS?顯影劑進行定量測定。

ENZO熱銷產(chǎn)品——FLUOR DE LYS? HDAC fluorometric activity assay kit組蛋白去乙?;笩晒饣钚詸z測試劑盒

Figure 1.?HDAC熒光活性檢測的反應方案。底物去乙?;蛊鋵︼@影劑敏感,從而產(chǎn)生熒光團(symbol)。熒光團可被360 nm的光激發(fā),并發(fā)射出460 nm的光,發(fā)射光在熒光平板閱讀器上可被檢測到。

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產(chǎn)品特點:

使用提供的核提取物,有助于檢測裂解物、免疫沉淀物以及篩選抑制劑;

含有HeLa核提取物,作為HDAC 1和2的豐富來源,可用作陽性對照,也可用作篩選HDAC活性的來源;

與I & IIb類HDAC和Sirtuin兼容(添加NAD+);

無放射性,沒有提取步驟;

所提供的試劑足夠分析100-200次。

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產(chǎn)品信息:

產(chǎn)品貨號BML-AK500-0001
產(chǎn)品名稱FLUOR DE LYS? HDAC fluorometric activity assay kit
規(guī)格1*96wells
使用/穩(wěn)定性Store all components, except the microtiter plate, at -80℃?for the highest stability. The HeLa Nuclear Extract, BML-KI140, must be handled with particular care in order to retain maximum enzymatic activity. Defrost it quickly in a RT water bath or by rubbing between fingers, then immediately store on an ice bath. The remaining unused extract should be refrozen quickly, by placing at -80°C. If possible, snap freeze in liquid nitrogen or a dry ice/ethanol bath. To minimize the number of freeze/thaw cycles, aliquot the extract into separate tubes and store at -80℃. The FLUOR DE LYS? Substrate, BML-KI104, when diluted in Assay Buffer, may precipitate after freezing and thawing. It is best, therefore, to dilute only the amount needed to perform the assays of that day.
運輸Dry Ice
長期保存-80℃
試劑盒組分Nuclear Extract from HeLa Cells?(human cervical cancer cell line) (Prod. No. BML-KI140-0100)
(100 μl; In 0.1M KCl, 20mM HEPES/NaOH, pH 7.9, 20% (v/v) glycerol, 0.2mM EDTA, 0.5mM DTT, 0.5Mm PMSF, prepared according to a modification of J.D. Dignam et al. (1983) and S.M. Abmayr et al. (1988)).
Storage: -80℃, avoid freeze/thaw cycles
FLUOR DE LYS? Substrate (Prod. No. BML-KI104-0050)
(50 μl; 50mM in DMSO)
Storage: -80℃
FLUOR DE LYS? Developer Concentrate (20×) (Prod. No. BML-KI105-0300) (300 μl; 20×stock solution, dilute in assay buffer before use)
Storage: -80℃
Trichostatin A (HDAC Inhibitor) (Prod. No. BML-GR309-9090)
(100 μl; 0.2mM in DMSO)
Storage: -80℃
FLUOR DE LYS? Deacetylated Standard (Prod. No. BML-KI142-0030)
(30 μl; 10mM in DMSO)
Storage: -80℃
HDAC Assay Buffer (Prod. No. BML-KI143-0020)
(20 ml; 50mM TRIS/Cl, pH 8.0, 137mM NaCl, 2.7mM KCl, 1mM MgCl2)
Storage: -20℃
1/2 volume microplate (Prod. No. BML-KI101)
Storage: Ambient
1/2 volume white microplate (Prod. No. BML-K571)
Storage: Ambient

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產(chǎn)品最新引用文獻:

The activation of histone deacetylases 4 prevented endothelial dysfunction: A crucial mechanism of HuangqiGuizhiWuwu Decoction in improving microcirculation dysfunction in diabetes: M. Chen, et al.; J. Ethnopharmacol.?307, 116240 (2023),?Abstract;

Four New Anthraquinones with Histone Deacetylase Inhibitory Activity from Ventilago denticulata Roots: N. Hangsamai, et al.; Molecules?27, 1088 (2022),?Abstract;

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